U.S. patent number 11,320,416 [Application Number 16/887,609] was granted by the patent office on 2022-05-03 for methods of assessing suitability of use of pharmaceutical compositions of albumin and poorly water soluble drug.
This patent grant is currently assigned to Abraxis BioScience, LLC. The grantee listed for this patent is Abraxis BioScience, LLC. Invention is credited to Neil P. Desai, Willard Foss, Viktor Peykov, Daniel W. Pierce.
United States Patent |
11,320,416 |
Peykov , et al. |
May 3, 2022 |
Methods of assessing suitability of use of pharmaceutical
compositions of albumin and poorly water soluble drug
Abstract
The present invention provides methods of assessing suitability
of a pharmaceutical composition for medical use. The pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin.
Inventors: |
Peykov; Viktor (San Diego,
CA), Foss; Willard (San Diego, CA), Pierce; Daniel W.
(Belmont, CA), Desai; Neil P. (Pacific Palisades, CA) |
Applicant: |
Name |
City |
State |
Country |
Type |
Abraxis BioScience, LLC |
Summit |
NJ |
US |
|
|
Assignee: |
Abraxis BioScience, LLC
(Summit, NJ)
|
Family
ID: |
1000005073522 |
Appl.
No.: |
16/887,609 |
Filed: |
May 29, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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16277265 |
Feb 15, 2019 |
10705070 |
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15928702 |
Mar 22, 2018 |
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15062048 |
Mar 5, 2016 |
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62129010 |
Mar 5, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
G01N
21/21 (20130101); G01N 21/35 (20130101); G01N
24/08 (20130101); G01N 23/20 (20130101); A61K
9/5169 (20130101); A61K 31/436 (20130101); G01N
33/4833 (20130101); G01N 2021/3595 (20130101) |
Current International
Class: |
G01N
33/483 (20060101); G01N 24/08 (20060101); A61K
9/51 (20060101); A61K 31/436 (20060101); G01N
21/35 (20140101); G01N 21/21 (20060101); G01N
23/20 (20180101) |
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|
Primary Examiner: Wax; Robert A
Assistant Examiner: Tcherkasskaya; Olga V.
Attorney, Agent or Firm: Morrison & Foerster LLP
Parent Case Text
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a continuation of U.S. patent application Ser.
No. 16/277,265 (now issued as U.S. Pat. No. 10,705,070), filed Feb.
15, 2019, which is a continuation of U.S. patent application Ser.
No. 15/928,702 (now abandoned), filed Mar. 22, 2018, which is a
continuation of U.S. patent application Ser. No. 15/062,048 (now
abandoned), filed Mar. 5, 2016, which claims priority of U.S.
Provisional Application No. 62/129,010, filed Mar. 5, 2015, each of
which is incorporated herein by reference in its entirety for all
purposes.
Claims
What is claimed is:
1. A method of validating a commercial batch of a pharmaceutical
composition for medical use in a human individual, wherein the
pharmaceutical composition comprises (a) nanoparticles comprising
rapamycin coated with a coating comprising albumin and (b) a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: (1) obtaining a sample from the commercial
batch; and (2) assessing suitability of the pharmaceutical
composition for use in a human individual, comprising: separating
the nanoparticles in the sample from the non-nanoparticle portion
of the sample; measuring a weight percentage of albumin in the form
of albumin polymers in the coating of the separated nanoparticles
among the total albumin in the coating of the separated
nanoparticles; and assessing the suitability of the pharmaceutical
composition for medical use in a human individual, wherein the
weight percentage of albumin in the form of albumin polymers in the
coating of the separated nanoparticles among the total albumin in
the coating of the separated nanoparticles being from about 15% to
about 40% is indicative of suitability of the pharmaceutical
composition for medical use; and (3) validating the commercial
batch if the pharmaceutical composition is suitable for medical
use.
2. The method of claim 1, wherein the method further comprises
measuring a weight percentage of albumin in the form of albumin
monomers in the coating of the separated nanoparticles among the
total albumin in the coating of the separated nanoparticles,
wherein weight percentage of albumin in the form of albumin
monomers among the total albumin in the coating of the separated
nanoparticles being from about 40% to about 60% is indicative of
suitability of the pharmaceutical composition for medical use.
3. The method of claim 1, wherein the method further comprises
determining a weight percentage of albumin in the coating of the
separated nanoparticles compared to the total weight of the
separated nanoparticles, wherein the weight percentage of the
albumin in the coating of the separated nanoparticles compared to
the total weight of the separated nanoparticles being from about
15% to about 30% is indicative of suitability of the pharmaceutical
composition for medical use.
4. The method of claim 1, further comprising determining a weight
ratio of albumin in the coating of the nanoparticles to rapamycin
in the nanoparticles, wherein a weight ratio of albumin in the
coating of the nanoparticles to rapamycin in the nanoparticles
being from about 1:2 to about 1:6 is indicative of suitability of
the pharmaceutical composition for medical use.
5. The method of claim 1, further comprising determining the
morphology of the nanoparticles by cryogenic transmission electron
microscopy, wherein an irregular shape of the nanoparticles is
indicative of suitability of the pharmaceutical composition for
medical use.
6. The method of claim 1, further comprising determining a
thickness of the coating of the nanoparticles by cryo transmission
electron microscopy, wherein the thickness being from about 5 nm to
about 7 nm is indicative of suitability of the pharmaceutical
composition for medical use.
7. The method of claim 1, further comprising determining a
solubility of the nanoparticles in a 5% human albumin solution by
dynamic light scattering, wherein the solubility being from about
50 .mu.g/ml to about 100 .mu.g/ml in the 5% human albumin solution
as determined by dynamic light scattering is indicative of
suitability of the pharmaceutical composition for medical use.
8. The method of claim 1, further comprising determining a
rapamycin crystallinity of the rapamycin in the nanoparticles of
the pharmaceutical composition, wherein the rapamycin being in a
non-crystalline state is indicative of suitability of the
pharmaceutical composition for medical use.
9. The method of claim 1, further comprising determining a
rapamycin recovery following a 0.2 micron filtration of the
pharmaceutical composition, wherein the rapamycin recovery being at
least about 80% is indicative of suitability of the pharmaceutical
composition for medical use.
10. The method of claim 9, wherein the determination of the
rapamycin recovery is carried out after storage.
11. The method of claim 8, wherein the rapamycin crystallinity is
determined by X-ray diffraction, polarized light microscopy, or
both.
12. The method of claim 1, further comprising determining a binding
affinity of albumin to rapamycin in the pharmaceutical
composition.
13. The method of claim 12, wherein the binding affinity is
determined by equilibrium dialysis, Fourier-transform infrared
spectroscopy, nuclear magnetic resonance, or a combination
thereof.
14. The method of claim 1, further comprising determining a
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition.
15. The method of claim 1, further comprising determining a weight
percentage of albumin in the form of albumin dimers in the coating
of the separated nanoparticles among the total albumin in the
coating of the separated nanoparticles, wherein the weight
percentage of albumin in the form of albumin dimers in the coating
of the separated nanoparticles among the total albumin in the
coating of the separated nanoparticles being from about 15% to
about 30% is indicative of the pharmaceutical composition being
suitable for medical use.
16. The method of claim 1, further comprising determining a weight
percentage of albumin in the form of albumin oligomers in the
coating of the separated nanoparticles among the total albumin in
the coating of the separated nanoparticles, wherein the weight
percentage of albumin in the form of albumin oligomers in the
coating of the separated nanoparticles among the total albumin in
the coating of the separated nanoparticles being from about 7% to
about 15% is indicative of the pharmaceutical composition being
suitable for medical use.
17. The method of claim 1, further comprising determining: a weight
percentage of albumin in the form of albumin monomers in the
pharmaceutical composition among the total albumin in the
pharmaceutical composition; a weight percentage of albumin in the
form of albumin dimers in the pharmaceutical composition among the
total albumin in the pharmaceutical composition; a weight
percentage of albumin in the form of albumin oligomers in the
pharmaceutical composition among the total albumin in the
pharmaceutical composition; or a weight percentage of albumin in
the form of albumin polymers in the pharmaceutical composition
among the total albumin in the pharmaceutical composition.
18. The method of claim 1, wherein the weight percentage of albumin
in the form of albumin monomers in the coating of the separated
nanoparticles among the total albumin in the coating of the
separated nanoparticles is measured by size-exclusion
chromatography.
19. The method of claim 1, further comprising determining a
particle size of the nanoparticles.
20. The method of claim 19, wherein the particle size of the
nanoparticles is determined by dynamic light scattering.
21. The method of claim 1, further comprising determining a
polydispersity index of the nanoparticles in the pharmaceutical
composition.
22. The method of claim 1, further comprising determining a span of
size distribution of the nanoparticles in the pharmaceutical
composition by (Dv.sub.90-Dv.sub.10)/Dv.sub.50; wherein: Dv.sub.90
is the particle diameter where 90% of the volume of all
nanoparticles is contained in nanoparticles with smaller diameters;
Dv.sub.10 is the particle diameter where 10% of the volume of all
nanoparticles is contained in nanoparticles with smaller diameters;
and Dv.sub.50 is the volume-weighted median particle diameter.
23. The method of claim 1, further comprising determining a surface
potential of the nanoparticles.
24. The method of claim 1, further comprising determining a weight
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition.
25. The method of claim 24, wherein the weight percentage of the
rapamycin in the nanoparticles among the total rapamycin in the
pharmaceutical composition is determined by reversed-phase
high-performance liquid chromatography.
26. The method of claim 1, further comprising determining a weight
percentage of albumin that is in the non-nanoparticle portion among
the total albumin in the pharmaceutical composition.
27. The method of claim 26, wherein the weight percentage of the
albumin that is in the non-nanoparticle portion among the total
albumin in the pharmaceutical composition is determined by
size-exclusion chromatography.
28. The method of claim 1, further comprising determining a
stability of the pharmaceutical composition.
29. The method of claim 28, wherein the stability is determined
after storage.
30. The method of claim 1, wherein a weight ratio of total albumin
in the pharmaceutical composition to total rapamycin in the
pharmaceutical composition is from about 3:1 to about 7.9:1 or from
about 10:1 to about 17:1.
31. The method of claim 1, wherein the albumin is human
albumin.
32. The method of claim 1, wherein the nanoparticles have a
volume-weighted median particle diameter of less than about 200
nm.
33. A commercial batch of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises (a) nanoparticles comprising rapamycin coated with a
coating comprising albumin and (b) a non-nanoparticle portion
comprising albumin and rapamycin, and wherein the commercial batch
is validated by assessment of suitability for medical use according
to claim 1.
34. The method of claim 1, further comprising determining a weight
percentage of albumin in the form of albumin monomers in the
coating of the separated nanoparticles among the total albumin in
the coating of the separated nanoparticles, wherein the weight
percentage of albumin in the form of albumin monomers among the
total albumin in the coating of the separated nanoparticles being
less than about 51% is indicative of the pharmaceutical composition
being suitable for medical use.
35. The method of claim 1, further comprising determining a ratio
of the sum of the weight percentage of albumin in the form of
albumin polymers in the coating of the separated nanoparticles and
a weight percentage of albumin in the form of oligomers in the
coating of the separated nanoparticles, compared to a weight
percentage of albumin in the form of albumin monomers in the
coating of the separated nanoparticles, wherein the ratio of the
sum of the weight percentage of albumin in the form of albumin
polymers in the coating of the separated nanoparticles and the
weight percentage of albumin in the form of oligomers in the
coating of the separated nanoparticles, compared to the weight
percentage of albumin in the form of albumin monomers in the
coating of the separated nanoparticles, being more than about 65%
is indicative of the pharmaceutical composition being suitable for
medical use.
36. The method of claim 17, comprising determining the weight
percentage of albumin in the form of albumin monomers in the
pharmaceutical composition among the total albumin in the
pharmaceutical composition; wherein the weight percentage of
albumin in the form of albumin monomers in the pharmaceutical
composition among the total albumin in the pharmaceutical
composition being from about 75% to about 87% is indicative of the
pharmaceutical composition being suitable for medical use.
37. The method of claim 17, comprising determining the weight
percentage of albumin in the form of albumin dimers in the
pharmaceutical composition among the total albumin in the
pharmaceutical composition; wherein the weight percentage of
albumin in the form of albumin dimers in the pharmaceutical
composition among the total albumin in the pharmaceutical
composition being from about 6% to about 13% is indicative of the
pharmaceutical composition being suitable for medical use.
38. The method of claim 17, comprising determining the weight
percentage of albumin in the form of albumin oligomers in the
pharmaceutical composition among the total albumin in the
pharmaceutical composition; wherein the weight percentage of
albumin in the form of albumin oligomers in the pharmaceutical
composition among the total albumin in the pharmaceutical
composition being from about 1% to about 4% is indicative of the
pharmaceutical composition being suitable for medical use.
39. The method of claim 17, comprising determining the weight
percentage of albumin in the form of albumin polymers in the
pharmaceutical composition among the total albumin in the
pharmaceutical composition; wherein the weight percentage of
albumin in the form of albumin polymers in the pharmaceutical
composition among the total albumin in the pharmaceutical
composition being at least about 1% is indicative of the
pharmaceutical composition being suitable for medical use.
40. The method of claim 17, comprising determining the weight
percentage of albumin in the form of albumin monomers in the
pharmaceutical composition among the total albumin in the
pharmaceutical composition and the weight percentage of albumin in
the form of albumin polymers in the pharmaceutical composition
among the total albumin in the pharmaceutical composition; wherein
the weight percentage of albumin in the form of albumin monomers in
the pharmaceutical composition among the total albumin in the
pharmaceutical composition being from about 75% to about 87% and
the weight percentage of albumin in the form of albumin polymers in
the pharmaceutical composition among the total albumin in the
pharmaceutical composition being from about 1% to about 5% is
indicative of the pharmaceutical composition being suitable for
medical use.
41. A method of releasing a commercial batch of a pharmaceutical
composition comprising (a) nanoparticles comprising rapamycin
coated with a coating comprising albumin and (b) a non-nanoparticle
portion comprising albumin and rapamycin, the method comprising:
validating the commercial batch according to the method of claim 1;
and releasing the commercial batch if the pharmaceutical
composition is suitable for medical use.
42. A method of processing a sample of a pharmaceutical composition
to assess suitability of the pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises (a) nanoparticles comprising rapamycin coated with a
coating comprising albumin and (b) a non-nanoparticle portion
comprising albumin and rapamycin, the method comprising: (1)
obtaining a sample from the commercial batch; and (2) processing
the sample to assess suitability of the pharmaceutical composition
for medical use in a human individual, comprising: separating the
nanoparticles in the sample from the non-nanoparticle portion of
the sample; measuring a weight percentage of albumin in the form of
albumin polymers in the coating of the separated nanoparticles
among the total albumin in the coating of the separated
nanoparticles; and assessing the suitability of the pharmaceutical
composition for medical use in a human individual, wherein the
weight percentage of albumin in the form of albumin polymers in the
coating of the separated nanoparticles among the total albumin in
the coating of the separated nanoparticles being from about 15% to
about 40% is indicative of suitability of the pharmaceutical
composition for medical use.
43. The commercial batch of claim 33, wherein the commercial batch
of the pharmaceutical composition is manufactured according to a
method comprising: homogenizing a mixture comprising a first
solution comprising an organic solvent and rapamycin and a second
solution comprising water and albumin to form a homogenized
mixture; and removing the organic solvent from the homogenized
mixture.
Description
TECHNICAL FIELD
The present invention relates to methods of assessing suitability
of use of a pharmaceutical composition of albumin and poorly water
soluble drug.
BACKGROUND
Albumin-based pharmaceutical compositions have been developed as a
drug delivery system for delivering a substantially water insoluble
drugs such as taxane. See, for example, U.S. Pat. Nos. 5,916,596 A,
6,506,405 B1, 6,749,868 B1, 6,537,579 B1, 7,820,788 B2, and
7,923,536 B2. For example, nab-paclitaxel sold under the trademark
ABRAXANE.RTM., an albumin-stabilized nanoparticle formulation of
paclitaxel, is a prescription drug approved to treat
life-threatening cancers that affect hundreds of thousands of
patients in the United States. Nab-paclitaxel sold under the
trademark ABRAXANE.RTM. is indicated for the treatment of
metastatic breast cancer, locally advanced or metastatic non-small
cell lung cancer ("NSCLC"), as well as metastatic adenocarcinoma of
the pancreas. Additionally, ABI-009, an albumin-stabilized
nanoparticle formulation of rapamycin, has been reported to exhibit
evidence of response in a Phase I trial of patients with advanced
nonhematologic malignancies. See Gonzalez-Angulo, A. M. et al.
Clin. Cancer Res. 2013.
It is generally believed that albumin-based nanoparticles, such as
those in ABI-009 and nab-paclitaxel sold under the trademark
ABRAXANE.RTM., when introduced into the blood stream, would
dissolve into albumin-drug complexes. Such albumin-drug complexes
utilize the natural properties of albumin to transport and deliver
substantially water insoluble drugs to the site of disease, such as
tumor sites. In addition, the albumin-based nanoparticle technology
offers the ability to improve a drug's solubility without the need
for toxic solvents in the administration process, thus potentially
improving safety through the elimination of solvent-related side
effects.
The disclosures of all publications, patents, patent applications,
and published patent applications referred to herein are hereby
incorporated herein by reference in their entireties.
BRIEF SUMMARY DESCRIBED HEREIN
The present application in some embodiment provides methods of
assessing suitability of a composition (such as a pharmaceutical
composition) for medical use, wherein the composition (such as a
pharmaceutical composition) comprises nanoparticles comprising
rapamycin coated with albumin and a non-nanoparticle portion
comprising albumin and a poorly water soluble drug (such as
rapamycin).
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
polymers among the albumin on the nanoparticles, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
percentage of albumin monomers among the albumin on the
nanoparticles, wherein a percentage of albumin monomers among the
albumin on the nanoparticles being about 40% to about 60% (such as
about 40% to about 55%, about 40% to about 54%, about 40% to about
53%, about 40% to about 52%, about 40% to about 50%, about 40% to
about 48%, or about 40% to about 46%) is indicative of suitability
of the pharmaceutical composition for medical use.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
monomers among the albumin on the nanoparticles, wherein a
percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments, a
percentage of albumin monomers among the albumin on the
nanoparticles being less than about 52% is indicative of
suitability of the pharmaceutical composition for medical use.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
polymers and oligomers among the albumin on the nanoparticles,
wherein a percentage of albumin polymers and oligomers among the
albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
polymers and monomers among the albumin on the nanoparticles,
wherein a percentage of albumin polymers among the albumin on the
nanoparticles being more than about 11% and a percentage of albumin
monomers among the albumin on the nanoparticles being less than
about 54% is indicative of suitability of the pharmaceutical
composition for medical use.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
polymers and monomers among the albumin on the nanoparticles,
wherein a percentage of albumin polymers among the albumin on the
nanoparticles being more than about 18% and a percentage of albumin
monomers among the albumin on the nanoparticles being less than
about 55% is indicative of suitability of the pharmaceutical
composition for medical use.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
polymers, oligomers, and monomers among the albumin on the
nanoparticles, wherein the ratio of albumin on the nanoparticles in
the forms of polymers and oligomers to the albumin on the
nanoparticles in the form of monomers being more than about 62%
indicative of suitability of the pharmaceutical composition for
medical use.
In some embodiments according to any of the methods described
above, the method further comprises determining the weight
percentage of the albumin in the nanoparticles, wherein a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%) is indicative of suitability of the
pharmaceutical composition for medical use.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the weight percentage of the
albumin in the nanoparticles, wherein a weight percentage of the
albumin in the nanoparticles being about 15% to about 30% (such as
about 20% to about 25%, about 15% to about 24%, or about 15% to
about 20%) is indicative of suitability of the pharmaceutical
composition for medical use.
In some embodiments according to any of the methods described
above, the method further comprises determining the weight ratio of
albumin to rapamycin in the nanoparticles, wherein an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles is
indicative of suitability of the pharmaceutical composition for
medical use.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles, wherein an albumin to rapamycin
ratio of about 1:2 to about 1:6 in the nanoparticles is indicative
of suitability of the pharmaceutical composition for medical
use.
In some embodiments according to any of the methods described
above, the method further comprises determining the morphology of
the nanoparticles under cryogenic transmission electron microscopy
(TEM), wherein an irregular shape of the nanoparticles is
indicative of suitability of the pharmaceutical composition for
medical use.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the morphology of the
nanoparticles under cryo-TEM, wherein an irregular shape of the
nanoparticles is indicative of suitability of the pharmaceutical
composition for medical use.
In some embodiments according to any of the methods described
above, the method further comprises determining the thickness of
the albumin coating of the nanoparticles under cryo-TEM, wherein a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the thickness of the albumin
coating of the nanoparticles under cryo-TEM, wherein a thickness of
about 5-7 nm (such as about 6 nm) is indicative of suitability of
the pharmaceutical composition for medical use.
In some embodiments according to any of the methods described
above, there is provided a method of assessing suitability of a
pharmaceutical composition for medical use in a human individual,
wherein the pharmaceutical composition comprises nanoparticles
comprising rapamycin coated with albumin and a non-nanoparticle
portion comprising albumin and rapamycin, the method comprising:
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue; wherein an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use.
In some embodiments according to any of the methods described
above, the method further comprises determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor; wherein an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use.
In some embodiments according to any of the methods described
above, the method further comprises determining the solubility of
the pharmaceutical composition, wherein a solubility of about 50
.mu.g/ml to about 100 .mu.g/ml in a 5% human albumin solution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the determination of solubility
is carried out after storage.
In some embodiments according to any of the methods described
above, the method further comprises determining the rapamycin
crystallinity of the pharmaceutical composition, wherein a
non-crystalline state of the rapamycin is indicative of suitability
of the pharmaceutical composition for medical use. In some
embodiments, the determination of rapamycin crystalline state is
carried out after storage. In some embodiments, the rapamycin
crystallinity is determined by X-ray diffraction, polarized light
microscopy, or both.
In some embodiments according to any of the methods described
above, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration of the pharmaceutical
composition, wherein a rapamycin recovery of at least about 80% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the determination of rapamycin
recovery is carried out after storage.
One aspect of the present application provides a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the solubility, rapamycin
crystallinity, and a rapamycin recovery following a 0.2 micron
filtration of the pharmaceutical composition, wherein a solubility
of about 50 .mu.g/ml to about 100 .mu.g/ml in a 5% human albumin
solution, a non-crystalline state of the rapamycin, and a rapamycin
recovery date of at least about 80% is indicative of suitability of
the pharmaceutical composition for medical use. In some
embodiments, the method is carried out after storage.
In some embodiments according to any of the methods described
above, the method further comprises determining the binding
affinity of albumin to rapamycin in the pharmaceutical composition.
In some embodiments, the binding affinity is determined by
equilibrium dialysis, Fourier-transform infrared spectroscopy
(FTIR), nuclear magnetic resonance (NMR), or a combination
thereof.
In some embodiments according to any of the methods described
above, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition.
In some embodiments according to any of the methods described
above, the method further comprises determining the percentage of
albumin dimers among the albumin on the nanoparticles, wherein a
percentage of about 15% to about 30% of albumin dimers among the
albumin on the nanoparticles is indicative of the pharmaceutical
composition for medical use.
In some embodiments according to any of the methods described
above, the method further comprises determining the percentage of
albumin oligomers among the albumin on the nanoparticles, wherein a
percentage of about 7% to about 15% of albumin oligomers among the
albumin on the nanoparticles is indicative of the pharmaceutical
composition for medical use.
In some embodiments according to any of the methods described
above, the method further comprises determining the percentage of
albumin monomers, dimers, oligomers, or polymers among the total
albumin in the pharmaceutical composition. In some embodiments, the
percentage of albumin monomers, dimers, oligomers, or polymers is
carried out by size-exclusion chromatography.
In some embodiments according to any of the methods described
above, the method further comprises determining the particle size
of the nanoparticles. In some embodiments, the particle size of the
nanoparticles is determined by dynamic light scattering.
In some embodiments according to any of the methods described
above, the method further comprises determining the polydispersity
index of the nanoparticles in the pharmaceutical composition.
In some embodiments according to any of the methods described
above, the method further comprises determining the span of size
distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles
in the pharmaceutical composition. Dv.sub.50 refers to the
volume-weighted median particle diameter. Dv.sub.90 refers to the
particle diameter where 90% of the volume of all nanoparticles is
contained in nanoparticles with smaller diameters. Dv.sub.10 refers
to the particle diameter where 10% of the volume of all
nanoparticles is contained in nanoparticles with smaller
diameters.
In some embodiments according to any of the methods described
above, the method further comprises determining the surface
potential of the nanoparticles.
In some embodiments according to any of the methods described
above, the method further comprises determining the percentage of
the rapamycin in the nanoparticles among the total rapamycin in the
pharmaceutical composition. In some embodiments, the percentage of
the rapamycin in the nanoparticles is determined by reversed-phase
high performance liquid chromatography (RP-HPLC).
In some embodiments according to any of the methods described
above, the method further comprises determining the percentage of
the albumin that is in the non-nanoparticle portion among the total
albumin in the pharmaceutical composition. In some embodiments, the
percentage of the albumin is determined by size-exclusion
chromatography.
In some embodiments according to any of the methods described
above, the method further comprises determining the stability of
the pharmaceutical composition. In some embodiments, the stability
is determined after storage.
In some embodiments according to any of the methods described
above, the method further comprises determining tumor distribution
of rapamycin upon administration in vivo.
In some embodiments, the method comprises determining tumor
distribution of rapamycin upon injection of the pharmaceutical
composition directly into the tumor tissue.
In some embodiments according to any of the methods described
above, the weight ratio of the total albumin to the total rapamycin
in the pharmaceutical composition is about 3:1 to about 7.9:1 or
about 10:1 to about 17:1.
In some embodiments according to any of the methods described
above, the albumin is human albumin.
In some embodiments according to any of the methods described
above, the average particle size of the nanoparticles is less than
about 200 nm (such as about 120 nm to about 140 nm, for example
about 130 nm).
In a further aspect of the present application, there is provided a
method of validating a commercial batch of a pharmaceutical
composition for medical use in a human individual, wherein the
pharmaceutical composition comprises nanoparticles comprising
rapamycin coated with albumin and a non-nanoparticle portion
comprising albumin and rapamycin, and wherein the method comprises
1) obtaining a sample from the commercial batch, and 2) assessing
suitability of the sample for medical use according to any one of
the methods of assessing as described above.
In a further aspect of the present application, there is provided a
commercial batch of a pharmaceutical composition for medical use in
a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
and wherein the commercial batch is validated by assessment of
suitability for medical use according to any one of the methods of
assessing as described above.
Also provided are kits, medicines, and articles of manufacture
comprising any one of the compositions (such as pharmaceutical
compositions) described above.
These and other aspects and advantages of the present invention
will become apparent from the subsequent detailed description and
the appended claims. It is to be understood that one, some, or all
of the properties of the various embodiments described herein may
be combined to form other embodiments of the present invention.
BRIEF DESCRIPTION OF THE FIGURES
FIG. 1A-C show representative imaging of pancreatic MIA PaCa-2
xenograft tumors 24 hours (FIG. 1A), 48 hours (FIG. 1B), and 72
hours (FIG. 1C) post-injection with nab-paclitaxel sold under the
trademark ABRAXANE.RTM. (ABX). Mitotically-arrested cells were
stained with an anti-pHH3 antibody (white).
FIG. 1D-F show representative imaging of pancreatic MIA PaCa-2
xenograft tumors 24 hours (FIG. 1D), 48 hours (FIG. 1E), and 72
hours (FIG. 1F) post-injection with a DMSO formulation of
paclitaxel (PTX:DMSO). Mitotically-arrested cells were stained with
an anti-pHH3 antibody (white).
FIG. 1G-I show representative imaging of pancreatic MIA PaCa-2
xenograft tumors 24 hours (FIG. 1G), 48 hours (FIG. 1H), and 72
hours (FIG. 1I) post-injection with a Cremophor EL formulation of
paclitaxel (PTX:CrEL). Mitotically-arrested cells were stained with
an anti-pHH3 antibody (white).
FIG. 2A-C show the fraction of pHH3 positive cells (fraction pHH3+)
versus the radial distance (.mu.m) as measured from the injection
site for pancreatic MIA PaCa-2 xenograft tumors 24 hours (FIG. 2A),
48 hours (FIG. 2B), and 72 hours (FIG. 2C) post-injection with
either ABX, PTX:DMSO, or PTX:CrEL.
FIG. 3A shows the fraction pHH3+ versus the radial distance (.mu.m)
as measured from the injection site for A2058 tumor xenografts 24
hours post-injection with either ABX, PTX:CrEL, CrEL, or PBS.
FIG. 3B shows the fraction pHH3+ versus the radial distance (.mu.m)
as measured from the injection site for H2122 tumor xenografts 24
hours post-injection with either ABX, PTX:DMSO, DMSO, or PBS.
FIG. 4A-C show the fraction pHH3+ versus the radial distance
(.mu.m) as measured from the injection site for pancreatic MIA
PaCa-2 xenograft tumors at 24 hours post-injection with either 1.6
mg/mL ABX, 1.6 mg/mL PTX:DMSO, or 1.6 mg/mL PTX:CrEL (FIG. 4A);
either 2.5 mg/mL ABX, 2.5 mg/mL PTX:DMSO, or 2.5 mg/mL PTX:CrEL
(FIG. 4B); and either 4.75 mg/mL ABX, 4.75 mg/mL PTX:DMSO, or 4.75
mg/mL PTX:CrEL (FIG. 4C). The level of background signal is
indicated with a dashed line.
FIG. 5 shows a chromatogram from the separation of polymeric,
oligomeric, dimeric, and monomeric albumin on nanoparticles from a
pharmaceutical composition using size-exclusion chromatography.
FIG. 6 shows a diagram of a UV-Vis spectrophotometer optical
system.
FIG. 7 shows in vitro dissolution kinetics of nab-paclitaxel sold
under the trademark ABRAXANE.RTM. in water at 100 .mu.g/ml
paclitaxel concentration, as measured at 340 nm by a UV-Vis
spectrophotometer with a 295 nm low wavelength cut-off filter.
FIG. 8 shows a bar graph of the albumin as a percentage of the
nanoparticle mass.
FIG. 9 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of monomers.
FIG. 10 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of dimers.
FIG. 11 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of oligomers.
FIG. 12 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of polymers.
FIG. 13 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of monomers (M) and dimers (D).
FIG. 14 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of monomers (M) minus the percentage of
albumin on the nanoparticles in the form of dimers (D).
FIG. 15 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of monomers (M) and oligomers (O).
FIG. 16 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of monomers (M) and polymers (P).
FIG. 17 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of monomers (M) minus the percentage of
albumin on the nanoparticles in the form of polymers (P).
FIG. 18 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of dimers (D) and oligomers (O).
FIG. 19 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of dimers (D) and polymers (P).
FIG. 20 shows a bar graph of the percentage of albumin on the
nanoparticles in the form of oligomers (O) and polymers (P).
FIG. 21 shows a bar graph of the ratio (reported as a percentage)
of the percentage of albumin on the nanoparticles in the form of
dimers (D) divided by the percentage of albumin on the
nanoparticles in the form of monomers (M).
FIG. 22 shows a bar graph of the ratio (reported as a percentage)
of the percentage of albumin on the nanoparticles in the form of
oligomers (O) divided by the percentage of albumin on the
nanoparticles in the form of monomers (M).
FIG. 23 shows a bar graph of the ratio (reported as a percentage)
of the percentage of albumin on the nanoparticles in the form of
polymers (P) divided by the percentage of albumin on the
nanoparticles in the form of monomers (M).
FIG. 24 shows a bar graph of the ratio (reported as a percentage)
of the percentage of albumin on the nanoparticles in the form of
polymers (P) and oligomers (O) divided by the percentage of albumin
on the nanoparticles in the form of monomers (M).
FIG. 25 shows a bar graph of the ratio (reported as a percentage)
of the percentage of albumin on the nanoparticles in the form of
polymers (P) and oligomers (O) divided by the percentage of albumin
on the nanoparticles in the form of monomers (M) minus dimers
(D).
DETAILED DESCRIPTION
The present application provides methods of assessing suitability
for medical use (for example, medical use in a human individual) of
an albumin-based nanoparticle composition (for example a
pharmaceutical composition) by determining one or a number of
physicochemical characteristics and functional attributes of the
composition. The pharmaceutical compositions comprise: a)
nanoparticles comprising a poorly water soluble drug (such as
rapamycin), coated with albumin, and b) a non-nanoparticle portion
comprising albumin and a poorly water soluble drug (such as
rapamycin). The methods comprise determination of at least one
(such as at least any of 2, 3, 4, 5, 6, 7, or 8) of the following
characteristics or attributes: i) the oligomeric status of the
albumin on the nanoparticles, including percentage of albumin
polymers and/or monomers on the nanoparticles; ii) the percent by
weight of the albumin in the nanoparticles; iii) the weight ratio
of the albumin to the poorly water soluble drug (such as rapamycin)
in the nanoparticles; iv) particle morphology, including shape,
thickness of the coating, or surface-to-volume ratio; v)
distribution of poorly water soluble drug (such as rapamycin) in a
tumor tissue upon administration of the composition; vi) particle
solubility; vii) poorly water soluble drug (such as rapamycin)
crystallinity; and viii) poorly water soluble drug (such as
rapamycin) recovery following a 0.2 micron filtration. The methods
may further comprise determination of at least one (such as at
least any of 2, 3, 4, 5, 6, 7, 8, or 9) of the following
characteristics or attributes: 1) binding affinity of albumin to
poorly water soluble drug (such as rapamycin) in the composition
(for example by equilibrium dialysis, FTIR, NMR, or a combination
thereof); 2) surface-to-volume ratio; 3) percentage of albumin
dimers and/or oligomers among the albumin on the nanoparticles; 4)
distribution of the total poorly water soluble drug (such as
rapamycin) and/or the total albumin between the nanoparticles and
the non-nanoparticle portion; 5) oligomeric status of the total
albumin in the composition; 6) particle size of the nanoparticles,
including average particle size, polydispersity, and/or size
distribution; 7) surface potential; 8) in vitro release kinetics;
and 9) physical stability.
The methods provided herein are useful, for example, for validating
and/or releasing a commercial batch of an albumin-based poorly
water soluble drug (such as rapamycin) nanoparticle
composition.
The compositions (such as pharmaceutical compositions) described
herein, once determined to be suitable for medical use in a human
individual, can be useful for treating various diseases, such as
cancer. The present application thus also provides compositions
(such as pharmaceutical compositions, including for example
commercial batches) determined to be suitable for medical use, as
well as methods of using such compositions (such as pharmaceutical
compositions) for the treatment of diseases, including cancer. Also
provided herein are kits, medicines, and dosage forms comprising
the compositions (such as pharmaceutical compositions) described
herein and for use in methods described herein.
The exemplary embodiments provided herein disclose pharmaceutical
compositions. It is to be understood that these are exemplary
compositions and that these descriptions apply equally to and
describe other compositions of the invention as provided herein,
such as compositions having any of the characteristics defined in
these exemplary embodiments.
The exemplary embodiments provided herein disclose nanoparticle
compositions comprising rapamycin. It is to be understood that
these are exemplary compositions and that these descriptions apply
equally to and describe other compositions of the invention as
provided herein, such as nanoparticle compositions comprising
poorly water soluble drugs (such as taxanes, 17-AAG, and
thiocolchicine dimer).
Definitions
The term "individual" refers to a mammal and includes, but is not
limited to, human, bovine, horse, feline, canine, rodent, or
primate.
It is understood that aspects and embodiments described herein
include "consisting" and/or "consisting essentially of" aspects and
embodiments.
Reference to "about" a value or parameter herein includes (and
describes) variations that are directed to that value or parameter
per se. For example, description referring to "about X" includes
description of "X."
The term "about X-Y" used herein has the same meaning as "about X
to about Y."
As used herein and in the appended claims, the singular forms "a,"
"or," and "the" include plural referents unless the context clearly
dictates otherwise.
"Monomers" used herein refers to a single albumin molecule without
intermolecular disulfide bonds.
"RRT" used herein refers to the retention time relative to the
albumin monomers retention time on a size-exclusion HPLC
chromatography.
"Dimers" used herein refers to albumin species having an RRT of
about 0.86 to about 0.97.
"Oligomers" used herein refers to albumin species having an RRT of
about 0.70 to about 0.85.
"Polymers" used herein refers to albumin species having an RRT of
about 0.57 to about 0.69.
"The total albumin" in a composition (such as a pharmaceutical
composition) comprises the albumin on the nanoparticles and the
albumin in the non-nanoparticle portion of the composition. "The
albumin on the nanoparticles" or "the albumin in the nanoparticles"
refers to the albumin coated on the poorly water soluble drug, such
as rapamycin, in the nanoparticles, or the albumin coating of the
nanoparticles. "The total poorly water soluble drug" in a
composition (such as a pharmaceutical composition) comprises the
poorly water soluble drug, such as rapamycin, in the nanoparticles
and the poorly water soluble drug, such as rapamycin, in the
non-nanoparticle portion of the composition.
"Weight percentage of albumin in the nanoparticles" used herein
refers to the weight percentage of albumin in the total weight of
the nanoparticles.
"Weight ratio of albumin to poorly water soluble drug in the
nanoparticles" used herein refers to the weight ratio of albumin on
the nanoparticles to the poorly water soluble drug, such as
rapamycin, on the nanoparticles.
Methods of Assessing Suitability of Albumin-Based Rapamycin
Nanoparticle Compositions for Medical Use
The present application provides a method of assessing suitability
of a composition (also referred to as "albumin-based nanoparticle
composition") for medical use in an individual, wherein the
composition comprises nanoparticles comprising a poorly water
soluble drug (such as rapamycin) coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin. The
methods comprise determination of at least one (such as at least
any of 2, 3, 4, 5, 6, 7, or 8) of the following characteristics or
attributes: i) the oligomeric status of the albumin on the
nanoparticles (i.e. the albumin coating), including percentage of
albumin polymers and/or monomers in the nanoparticles; ii) the
percent by weight of the albumin in the nanoparticles; iii) the
weight ratio of the albumin to the poorly water soluble drug (such
as rapamycin) in the nanoparticles; iv) particle morphology,
including shape, thickness of the coating, and surface-to-volume
ratio; v) distribution of poorly water soluble drug (such as
rapamycin) in a tumor tissue upon administration of the composition
(for example upon direct injection of the composition directly into
the tumor tissue); vi) particle solubility; vii) poorly water
soluble drug (such as rapamycin) crystallinity; and viii) poorly
water soluble drug (such as rapamycin) recovery following a 0.2
micron filtration. The methods may further comprise determination
of at least one (such as at least any of 2, 3, 4, 5, 6, 7, 8, 9, or
10) of the following characteristics or attributes: 1) binding
affinity of albumin to poorly water soluble drug (such as
rapamycin) in the composition (for example by equilibrium dialysis,
FTIR, NMR, or a combination thereof); 2) surface-to-volume ratio;
3) percentage of albumin dimers and/or oligomers among the albumin
on the nanoparticles; 4) distribution of the total poorly water
soluble drug (such as rapamycin) and/or the total albumin between
the nanoparticles and the non-nanoparticle portion; 5) oligomeric
status of the total albumin in the composition; 6) particle size of
the nanoparticles, including average particle size, polydispersity,
and/or size distribution; 7) surface potential; 8) in vitro release
kinetics; 9) physical stability; and, in some embodiments, 10)
poorly water soluble drug (such as rapamycin) tumor distribution in
vivo.
Unless otherwise indicated, discussion of a certain parameter as
being indicative of suitability for medical use suggest that such
parameter may be determined in the method described herein. The
method thus, in some embodiments, encompasses a step of determining
such a parameter.
The compositions (such as pharmaceutical compositions) described
herein can be in liquid or powder forms. For example, in some
embodiments, the composition is a liquid nanoparticle suspension
(for example prior to lyophilization). In some embodiments, the
composition is a reconstituted suspension (e.g., in an aqueous
solution such as a saline solution). In some embodiments, the
poorly water soluble drug (such as rapamycin) concentration in the
suspension is about any of 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6
mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, or 10 mg/ml. In some embodiments,
the poorly water soluble drug (such as rapamycin) in the suspension
is about 5 mg/ml. In some embodiments, the composition is
lyophilized. In some embodiments, the composition is sterile. In
some embodiments, the composition is contained in a sealed
vial.
Thus, in some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments, a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the size of the nanoparticles (for example by
dynamic light scattering). In some embodiments, the method further
comprises determining the polydispersity index of the nanoparticles
in the pharmaceutical composition. In some embodiments, the method
further comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, wherein a percentage of
albumin monomers among the albumin on the nanoparticles being about
40% to about 60% (such as about 40% to about 55%, about 40% to
about 54%, about 40% to about 53%, about 40% to about 52%, about
40% to about 50%, about 40% to about 48%, or about 40% to about
46%) is indicative of suitability of the pharmaceutical composition
for medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as any of about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), and a
percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, wherein a weight percentage of the albumin in
the nanoparticles being about 15% to about 30% (such as about 20%
to about 25%, about 15% to about 24%, or about 15% to about 20%) is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles and determining the weight
percentage of the albumin in the nanoparticles, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%) and a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles and determining the weight
percentage of the albumin in the nanoparticles, wherein a
percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%) and a weight percentage of the albumin
in the nanoparticles being about 15% to about 30% (such as about
20% to about 25%, about 15% to about 24%, or about 15% to about
20%) is indicative of suitability of the pharmaceutical composition
for medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles and determining
the weight percentage of the albumin in the nanoparticles, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), and a weight percentage of the albumin in the
nanoparticles being about 15% to about 30% (such as about 20% to
about 25%, about 15% to about 24%, or about 15% to about 20%) is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles, wherein an albumin to rapamycin
ratio of about 1:2 to about 1:6 in the nanoparticles is indicative
of suitability of the pharmaceutical composition for medical use.
In some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles and determining the weight
ratio of albumin to rapamycin in the nanoparticles, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%) and an albumin
to rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles and determining the weight
ratio of albumin to rapamycin in the nanoparticles, wherein a
percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), and an albumin to rapamycin ratio of
about 1:2 to about 1:6 in the nanoparticles is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles and determining
the weight ratio of albumin to rapamycin in the nanoparticles,
wherein a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as any of about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), and a
percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), and an albumin to rapamycin ratio
of about 1:2 to about 1:6 in the nanoparticles is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, and determining the weight ratio of albumin
to rapamycin in the nanoparticles, wherein a weight percentage of
the albumin in the nanoparticles being about 15% to about 30% (such
as about 20% to about 25%, about 15% to about 24%, or about 15% to
about 20%) and an albumin to rapamycin ratio of about 1:2 to about
1:6 in the nanoparticles is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
weight ratio of albumin to rapamycin in the nanoparticles, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%) and an albumin to rapamycin ratio of
about 1:2 to about 1:6 in the nanoparticles is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
weight ratio of albumin to rapamycin in the nanoparticles, wherein
a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), a weight percentage of the albumin in
the nanoparticles being about 15% to about 30% (such as about 20%
to about 25%, about 15% to about 24%, or about 15% to about 20%)
and an albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the method
further comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles, and
determining the weight ratio of albumin to rapamycin in the
nanoparticles, wherein a percentage of albumin polymer among the
albumin on the nanoparticles being about 15% to about 40% (such as
about 15% to about 20%, about 20% to about 24.5%, about 24.5% to
about 30%, about 30% to about 35%, or about 35% to about 40%), a
percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), a weight percentage of the albumin
in the nanoparticles being about 15% to about 30% (such as about
20% to about 25%, about 15% to about 24%, or about 15% to about
20%), and an albumin to rapamycin ratio of about 1:2 to about 1:6
in the nanoparticles is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the morphology of the nanoparticles
under cryo-TEM, wherein an irregular shape of the nanoparticles is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles and determining the
morphology of the nanoparticles under cryo-TEM, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%) and an irregular
shape of the nanoparticles is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles and determining the
morphology of the nanoparticles under cryo-TEM, wherein a
percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%) and about 40% to about 60% (such as
about 40% to about 55%, about 40% to about 54%, about 40% to about
53%, about 40% to about 52%, about 40% to about 50%, about 40% to
about 48%, or about 40% to about 46%) and an irregular shape of the
nanoparticles is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the method
further comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles and determining
the morphology of the nanoparticles under cryo-TEM, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), and an irregular shape of the nanoparticles is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, and determining the morphology of the
nanoparticles under cryo-TEM, wherein a weight percentage of the
albumin in the nanoparticles being about 15% to about 30% (such as
about 20% to about 25%, about 15% to about 24%, or about 15% to
about 20%) and an irregular shape of the nanoparticles is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
morphology of the nanoparticles under cryo-TEM, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), and an irregular shape of the
nanoparticles is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the method
further comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
morphology of the nanoparticles under cryo-TEM, wherein a
percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), a weight percentage of the albumin in
the nanoparticles being about 15% to about 30% (such as about 20%
to about 25%, about 15% to about 24%, or about 15% to about 20%),
and an irregular shape of the nanoparticles is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles, and
determining the morphology of the nanoparticles under cryo-TEM,
wherein a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), a weight percentage of the albumin in the nanoparticles
being about 15% to about 30% (such as about 20% to about 25%, about
15% to about 24%, or about 15% to about 20%), and an irregular
shape of the nanoparticles is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles and determining the morphology of
the nanoparticles under cryo-TEM, wherein an albumin to rapamycin
ratio of about 1:2 to about 1:6 in the nanoparticles and an
irregular shape of the nanoparticles is indicative of suitability
of the pharmaceutical composition for medical use. In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, and determining
the morphology of the nanoparticles under cryo-TEM, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles, and
an irregular shape of the nanoparticles is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, and determining
the morphology of the nanoparticles under cryo-TEM, wherein a
percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles and an irregular shape of the
nanoparticles is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the method
further comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight ratio of albumin to rapamycin in the nanoparticles, and
determining the morphology of the nanoparticles under cryo-TEM,
wherein a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), an albumin to rapamycin ratio of about 1:2 to about 1:6
in the nanoparticles, and an irregular shape of the nanoparticles
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the weight ratio of albumin to
rapamycin in the nanoparticles, and determining the morphology of
the nanoparticles under cryo-TEM, wherein a weight percentage of
the albumin in the nanoparticles being about 15% to about 30% (such
as about 20% to about 25%, about 15% to about 24%, or about 15% to
about 20%), an albumin to rapamycin ratio of about 1:2 to about 1:6
in the nanoparticles, and an irregular shape of the nanoparticles
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles, and
determining the morphology of the nanoparticles under cryo-TEM,
wherein a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, and an irregular shape of
the nanoparticles is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles, and
determining the morphology of the nanoparticles under cryo-TEM,
wherein a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), a weight percentage of the albumin in
the nanoparticles being about 15% to about 30% (such as about 20%
to about 25%, about 15% to about 24%, or about 15% to about 20%),
an albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles, and an irregular shape of the nanoparticles is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the weight ratio of albumin to rapamycin in the
nanoparticles, and determining the morphology of the nanoparticles
under cryo-TEM, wherein a percentage of albumin polymer among the
albumin on the nanoparticles being about 15% to about 40% (such as
about 15% to about 20%, about 20% to about 24.5%, about 24.5% to
about 30%, about 30% to about 35%, or about 35% to about 40%), a
percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), a weight percentage of the albumin
in the nanoparticles being about 15% to about 30% (such as about
20% to about 25%, about 15% to about 24%, or about 15% to about
20%), an albumin to rapamycin ratio of about 1:2 to about 1:6 in
the nanoparticles, and an irregular shape of the nanoparticles is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the thickness of the albumin coating
of the nanoparticles under cryo-TEM, wherein a thickness of about
5-7 nm (such as about 6 nm) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles and determining the
thickness of the albumin coating of the nanoparticles under
cryo-TEM, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%) and a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles and determining the
thickness of the albumin coating of the nanoparticles under
cryo-TEM, wherein a percentage of albumin monomers among the
albumin on the nanoparticles being about 40% to about 60% (such as
about 40% to about 55%, about 40% to about 54%, about 40% to about
53%, about 40% to about 52%, about 40% to about 50%, about 40% to
about 48%, or about 40% to about 46%) and a thickness of about 5-7
nm (such as about 6 nm) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles and determining
the thickness of the albumin coating of the nanoparticles under
cryo-TEM, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), a
percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), and a thickness of about 5-7 nm
(such as about 6 nm) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, and determining the thickness of the albumin
coating of the nanoparticles under cryo-TEM, wherein a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%) and a thickness of about 5-7 nm (such as
about 6 nm) is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the method
further comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
thickness of the albumin coating of the nanoparticles under
cryo-TEM, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), and a thickness of about 5-7 nm (such
as about 6 nm) is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the method
further comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
thickness of the albumin coating of the nanoparticles under
cryo-TEM, wherein a percentage of albumin monomers among the
albumin on the nanoparticles being about 40% to about 60% (such as
about 40% to about 55%, about 40% to about 54%, about 40% to about
53%, about 40% to about 52%, about 40% to about 50%, about 40% to
about 48%, or about 40% to about 46%), a weight percentage of the
albumin in the nanoparticles being about 15% to about 30% (such as
about 20% to about 25%, about 15% to about 24%, or about 15% to
about 20%), and a thickness of about 5-7 nm (such as about 6 nm) is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles, and
determining the thickness of the albumin coating of the
nanoparticles under cryo-TEM, wherein a percentage of albumin
polymer among the albumin on the nanoparticles being about 15% to
about 40% (such as about 15% to about 20%, about 20% to about
24.5%, about 24.5% to about 30%, about 30% to about 35%, or about
35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), a weight percentage
of the albumin in the nanoparticles being about 15% to about 30%
(such as about 20% to about 25%, about 15% to about 24%, or about
15% to about 20%), and a thickness of about 5-7 nm (such as about 6
nm) is indicative of suitability of the pharmaceutical composition
for medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles, and determining the thickness of
the albumin coating of the nanoparticles under cryo-TEM, wherein an
albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles, and a thickness of about 5-7 nm (such as about 6 nm)
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, and determining
the thickness of the albumin coating of the nanoparticles under
cryo-TEM, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), an albumin
to rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles,
and a thickness of about 5-7 nm (such as about 6 nm) is indicative
of suitability of the pharmaceutical composition for medical use.
In some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, and determining
the thickness of the albumin coating of the nanoparticles under
cryo-TEM, wherein a percentage of albumin monomers among the
albumin on the nanoparticles being about 40% to about 60% (such as
about 40% to about 55%, about 40% to about 54%, about 40% to about
53%, about 40% to about 52%, about 40% to about 50%, about 40% to
about 48%, or about 40% to about 46%), an albumin to rapamycin
ratio of about 1:2 to about 1:6 in the nanoparticles, and a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight ratio of albumin to rapamycin in the nanoparticles and
determining the thickness of the albumin coating of the
nanoparticles under cryo-TEM, wherein a percentage of albumin
polymer among the albumin on the nanoparticles being about 15% to
about 40% (such as any of about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%), and a percentage of albumin monomer among
the albumin on the nanoparticles being at least about 40% to about
60% (such as about 40% to about 55%, about 40% to about 54%, about
40% to about 53%, about 40% to about 52%, about 40% to about 50%,
about 40% to about 48%, or about 40% to about 46%), an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles, and
a thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the weight ratio of albumin to
rapamycin in the nanoparticles, and determining the thickness of
the albumin coating of the nanoparticles under cryo-TEM, wherein a
weight percentage of the albumin in the nanoparticles being about
15% to about 30% (such as about 20% to about 25%, about 15% to
about 24%, or about 15% to about 20%), an albumin to rapamycin
ratio of about 1:2 to about 1:6 in the nanoparticles, and a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles, and
determining the thickness of the albumin coating of the
nanoparticles under cryo-TEM, wherein a percentage of albumin
polymer among the albumin on the nanoparticles being about 15% to
about 40% (such as about 15% to about 20%, about 20% to about
24.5%, about 24.5% to about 30%, about 30% to about 35%, or about
35% to about 40%), a weight percentage of the albumin in the
nanoparticles being about 15% to about 30% (such as about 20% to
about 25%, about 15% to about 24%, or about 15% to about 20%), an
albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles, and a thickness of about 5-7 nm (such as about 6 nm)
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles, and
determining the thickness of the albumin coating of the
nanoparticles under cryo-TEM, wherein a percentage of albumin
monomers among the albumin on the nanoparticles being about 40% to
about 60% (such as about 40% to about 55%, about 40% to about 54%,
about 40% to about 53%, about 40% to about 52%, about 40% to about
50%, about 40% to about 48%, or about 40% to about 46%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, and a thickness of about 5-7
nm (such as about 6 nm) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the weight ratio of albumin to rapamycin in the
nanoparticles, and determining the thickness of the albumin coating
of the nanoparticles under cryo-TEM, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), and a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, and a thickness of about 5-7
nm (such as about 6 nm) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the morphology and thickness of the
albumin coating of the nanoparticles under cryo-TEM, wherein an
irregular shape of the nanoparticles and a thickness of about 5-7
nm (such as about 6 nm) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles and determining the
morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, wherein a percentage of albumin polymer
among the albumin on the nanoparticles being about 15% to about 40%
(such as about 15% to about 20%, about 20% to about 24.5%, about
24.5% to about 30%, about 30% to about 35%, or about 35% to about
40%), and an irregular shape of the nanoparticles and a thickness
of about 5-7 nm (such as about 6 nm) is indicative of suitability
of the pharmaceutical composition for medical use. In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles and determining the
morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, wherein a percentage of albumin monomers
among the albumin on the nanoparticles being about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), an irregular shape of
the nanoparticles, and a thickness of about 5-7 nm (such as about 6
nm) is indicative of suitability of the pharmaceutical composition
for medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles and determining
the morphology of the nanoparticles and thickness of the
nanoparticles under cryo-TEM, wherein a percentage of albumin
polymer among the albumin on the nanoparticles being about 15% to
about 40% (such as about 15% to about 20%, about 20% to about
24.5%, about 24.5% to about 30%, about 30% to about 35%, or about
35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), an irregular shape of
the nanoparticles, and a thickness of about 5-7 nm (such as about 6
nm) is indicative of suitability of the pharmaceutical composition
for medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, and determining the morphology of the
nanoparticles and thickness of the nanoparticles under cryo-TEM,
wherein a weight percentage of the albumin in the nanoparticles
being about 15% to about 30% (such as about 20% to about 25%, about
15% to about 24%, or about 15% to about 20%), an irregular shape of
the nanoparticles, and a thickness of about 5-7 nm (such as about 6
nm) is indicative of suitability of the pharmaceutical composition
for medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, wherein a percentage of albumin polymer
among the albumin on the nanoparticles being about 15% to about 40%
(such as about 15% to about 20%, about 20% to about 24.5%, about
24.5% to about 30%, about 30% to about 35%, or about 35% to about
40%), a weight percentage of the albumin in the nanoparticles being
about 15% to about 30% (such as about 20% to about 25%, about 15%
to about 24%, or about 15% to about 20%), an irregular shape of the
nanoparticles, and a thickness of about 5-7 nm (such as about 6 nm)
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, wherein a percentage of albumin monomers
among the albumin on the nanoparticles being about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), a weight percentage
of the albumin in the nanoparticles being about 15% to about 30%
(such as about 20% to about 25%, about 15% to about 24%, or about
15% to about 20%), an irregular shape of the nanoparticles, and a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles, and
determining the morphology of the nanoparticles and thickness of
the albumin coating under cryo-TEM, wherein a percentage of albumin
polymer among the albumin on the nanoparticles being about 15% to
about 40% (such as about 15% to about 20%, about 20% to about
24.5%, about 24.5% to about 30%, about 30% to about 35%, or about
35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), a weight percentage
of the albumin in the nanoparticles being about 15% to about 30%
(such as about 20% to about 25%, about 15% to about 24%, or about
15% to about 20%), an irregular shape of the nanoparticles, and a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles, determining the morphology of the
nanoparticles and thickness of the albumin coating under cryo-TEM,
wherein an albumin to rapamycin ratio of about 1:2 to about 1:6 in
the nanoparticles, an irregular shape of the nanoparticles, and a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, and determining
the morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, wherein a percentage of albumin polymer
among the albumin on the nanoparticles being about 15% to about 40%
(such as about 15% to about 20%, about 20% to about 24.5%, about
24.5% to about 30%, about 30% to about 35%, or about 35% to about
40%), an albumin to rapamycin ratio of about 1:2 to about 1:6 in
the nanoparticles, an irregular shape of the nanoparticles, and a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, and determining
the morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, wherein a percentage of albumin monomers
among the albumin on the nanoparticles being about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles, an
irregular shape of the nanoparticles, and a thickness of about 5-7
nm (such as about 6 nm) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight ratio of albumin to rapamycin in the nanoparticles, and
determining the morphology of the nanoparticles and thickness of
the albumin coating under cryo-TEM, wherein a percentage of albumin
polymer among the albumin on the nanoparticles being about 15% to
about 40% (such as about 15% to about 20%, about 20% to about
24.5%, about 24.5% to about 30%, about 30% to about 35%, or about
35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles, an
irregular shape of the nanoparticles, and a thickness of about 5-7
nm (such as about 6 nm) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the weight ratio of albumin to
rapamycin in the nanoparticles, and determining the morphology of
the nanoparticles and thickness of the albumin coating under
cryo-TEM, wherein a weight percentage of the albumin in the
nanoparticles being about 15% to about 30% (such as about 20% to
about 25%, about 15% to about 24%, or about 15% to about 20%), an
albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles, an irregular shape of the nanoparticles, and a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles, and
determining the morphology of the nanoparticles and thickness of
the albumin coating under cryo-TEM, wherein a percentage of albumin
polymer among the albumin on the nanoparticles being about 15% to
about 40% (such as about 15% to about 20%, about 20% to about
24.5%, about 24.5% to about 30%, about 30% to about 35%, or about
35% to about 40%), a weight percentage of the albumin in the
nanoparticles being about 15% to about 30% (such as about 20% to
about 25%, about 15% to about 24%, or about 15% to about 20%), an
albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles, an irregular shape of the nanoparticles, and a
thickness of about 5-7 nm (such as about 6 nm) is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles, and
determining the morphology of the nanoparticles and thickness of
the albumin coating under cryo-TEM, wherein a percentage of albumin
monomers among the albumin on the nanoparticles being about 40% to
about 60% (such as about 40% to about 55%, about 40% to about 54%,
about 40% to about 53%, about 40% to about 52%, about 40% to about
50%, about 40% to about 48%, or about 40% to about 46%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, an irregular shape of the
nanoparticles, and a thickness of about 5-7 nm (such as about 6 nm)
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the method further comprises
determining the solubility of the pharmaceutical composition
(including determining solubility after storage). In some
embodiments, a percentage of albumin monomer among the albumin on
the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the weight ratio of albumin to rapamycin in the
nanoparticles, and determining the morphology of the nanoparticles
and thickness of the albumin coating under cryo-TEM, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), a weight percentage of the albumin in the nanoparticles
being about 15% to about 30% (such as about 20% to about 25%, about
15% to about 24%, or about 15% to about 20%), an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles, an
irregular shape of the nanoparticles, and a thickness of about 5-7
nm (such as about 6 nm) is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the method further comprises determining the solubility of the
pharmaceutical composition (including determining solubility after
storage). In some embodiments, a percentage of albumin monomer
among the albumin on the nanoparticles being less than about 51% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
and oligomers among the albumin on the nanoparticles being more
than about 35% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers among the albumin on the nanoparticles being more
than about 17% and a percentage of albumin monomer among the
albumin on the nanoparticles being less than about 54% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 18%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the distribution of rapamycin in a
tumor tissue upon injection of the pharmaceutical composition
directly into the tumor tissue; wherein an enhanced rapamycin tumor
distribution is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if upon tumor injection it allows rapamycin to spread radially for
a distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%) and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%) and an enhanced rapamycin tumor
distribution is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if upon tumor injection it allows rapamycin to spread radially for
a distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles and determining
the distribution of rapamycin in a tumor tissue upon injection of
the pharmaceutical composition directly into the tumor tissue,
wherein a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, and determining the distribution of rapamycin
in a tumor tissue upon injection of the pharmaceutical composition
directly into the tumor tissue, wherein a weight percentage of the
albumin in the nanoparticles being about 15% to about 30% (such as
about 20% to about 25%, about 15% to about 24%, or about 15% to
about 20%) and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), and an enhanced rapamycin tumor
distribution is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if upon tumor injection it allows rapamycin to spread radially for
a distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), a weight percentage of the albumin in
the nanoparticles being about 15% to about 30% (such as about 20%
to about 25%, about 15% to about 24%, or about 15% to about 20%)
and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), a
percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), a weight percentage of the albumin
in the nanoparticles being about 15% to about 30% (such as about
20% to about 25%, about 15% to about 24%, or about 15% to about
20%), and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles and
an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, and determining
the distribution of rapamycin in a tumor tissue upon injection of
the pharmaceutical composition directly into the tumor tissue,
wherein a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles and
an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, and determining
the distribution of rapamycin in a tumor tissue upon injection of
the pharmaceutical composition directly into the tumor tissue,
wherein a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, and an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight ratio of albumin to rapamycin in the nanoparticles, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), a
percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), an albumin to rapamycin ratio of
about 1:2 to about 1:6, and an enhanced rapamycin tumor
distribution is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if upon tumor injection it allows rapamycin to spread radially for
a distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the weight ratio of albumin to
rapamycin in the nanoparticles, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, and an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, and an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin monomers among the albumin
on the nanoparticles being about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), a weight percentage of the albumin
in the nanoparticles being about 15% to about 30% (such as about
20% to about 25%, about 15% to about 24%, or about 15% to about
20%), an albumin to rapamycin ratio of about 1:2 to about 1:6 in
the nanoparticles, and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the weight ratio of albumin to rapamycin in the
nanoparticles, and determining the distribution of rapamycin in a
tumor tissue upon injection of the pharmaceutical composition
directly into the tumor tissue, wherein a percentage of albumin
polymer among the albumin on the nanoparticles being about 15% to
about 40% (such as about 15% to about 20%, about 20% to about
24.5%, about 24.5% to about 30%, about 30% to about 35%, or about
35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), a weight percentage
of the albumin in the nanoparticles being about 15% to about 30%
(such as about 20% to about 25%, about 15% to about 24%, or about
15% to about 20%), an albumin to rapamycin ratio of about 1:2 to
about 1:6 in the nanoparticles, and an enhanced rapamycin tumor
distribution is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the method
further comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the rapamycin
crystallinity of the pharmaceutical composition (for example by
X-ray diffraction and/or polarized light microscopy, including
determining crystallinity after storage). In some embodiments, the
method further comprises determining the rapamycin recovery
following a 0.2 micron filtration (including determining recovery
after storage). In some embodiments, the method further comprises
determining binding affinity of albumin to rapamycin in the
composition (such as a pharmaceutical composition) (for example by
equilibrium dialysis, FTIR, NMR, or a combination thereof). In some
embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the morphology of the nanoparticles
under cryo-TEM and determining the distribution of rapamycin in a
tumor tissue upon injection of the pharmaceutical composition
directly into the tumor tissue, wherein an irregular shape of the
nanoparticles and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the morphology
of the nanoparticles under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), an irregular
shape of the nanoparticles, and an enhanced rapamycin tumor
distribution is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if upon tumor injection it allows rapamycin to spread radially for
a distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the morphology
of the nanoparticles under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), an irregular shape of the
nanoparticles, and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the morphology of the nanoparticles under cryo-TEM, and determining
the distribution of rapamycin in a tumor tissue upon injection of
the pharmaceutical composition directly into the tumor tissue,
wherein a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), an irregular shape of the nanoparticles, and an
enhanced rapamycin tumor distribution is indicative of suitability
of the pharmaceutical composition for medical use. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if upon tumor injection it allows
rapamycin to spread radially for a distance that is greater than
(for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the morphology of the
nanoparticles under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an irregular shape of the
nanoparticles, and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
morphology of the nanoparticles under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an irregular shape of the
nanoparticles, and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
morphology of the nanoparticles under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), a weight percentage of the albumin in
the nanoparticles being about 15% to about 30% (such as about 20%
to about 25%, about 15% to about 24%, or about 15% to about 20%),
an irregular shape of the nanoparticles, and an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the morphology of the nanoparticles under cryo-TEM, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), a
percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), a weight percentage of the albumin
in the nanoparticles being about 15% to about 30% (such as about
20% to about 25%, about 15% to about 24%, or about 15% to about
20%), an irregular shape of the nanoparticles, and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles, determining the morphology of the
nanoparticles under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles, an
irregular shape of the nanoparticles, and an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, determining the
morphology of the nanoparticles under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles, an
irregular shape of the nanoparticles, and an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, determining the
morphology of the nanoparticles under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, an irregular shape of the
nanoparticles, and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight ratio of albumin to rapamycin in the nanoparticles,
determining the morphology of the nanoparticles under cryo-TEM, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), a
percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), an albumin to rapamycin ratio of
about 1:2 to about 1:6 in the nanoparticles, an irregular shape of
the nanoparticles, and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the weight ratio of albumin to
rapamycin in the nanoparticles, determining the morphology of the
nanoparticles under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, an irregular shape of the
nanoparticles, and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles,
determining the morphology of the nanoparticles under cryo-TEM, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, irregular shape of the
nanoparticles, and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles,
determining the morphology of the nanoparticles under cryo-TEM, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin monomers among the albumin
on the nanoparticles being about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), a weight percentage of the albumin
in the nanoparticles being about 15% to about 30% (such as about
20% to about 25%, about 15% to about 24%, or about 15% to about
20%), an albumin to rapamycin ratio of about 1:2 to about 1:6 in
the nanoparticles, an irregular shape of the nanoparticles, and an
enhanced rapamycin tumor distribution is indicative of suitability
of the pharmaceutical composition for medical use. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if upon tumor injection it allows
rapamycin to spread radially for a distance that is greater than
(for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the weight ratio of albumin to rapamycin in the
nanoparticles, determining the morphology of the nanoparticles
under cryo-TEM, and determining the distribution of rapamycin in a
tumor tissue upon injection of the pharmaceutical composition
directly into the tumor tissue, wherein a percentage of albumin
polymer among the albumin on the nanoparticles being about 15% to
about 40% (such as about 15% to about 20%, about 20% to about
24.5%, about 24.5% to about 30%, about 30% to about 35%, or about
35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), a weight percentage
of the albumin in the nanoparticles being about 15% to about 30%
(such as about 20% to about 25%, about 15% to about 24%, or about
15% to about 20%), an albumin to rapamycin ratio of about 1:2 to
about 1:6 in the nanoparticles, an irregular shape of the
nanoparticles, and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the thickness of the albumin coating
of the nanoparticles under cryo-TEM and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a thickness of about 5-7 nm (such as about 6 nm) and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the thickness
of the albumin coating of the nanoparticles under cryo-TEM, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin polymer among the albumin
on the nanoparticles being about 15% to about 40% (such as about
15% to about 20%, about 20% to about 24.5%, about 24.5% to about
30%, about 30% to about 35%, or about 35% to about 40%), a
thickness of about 5-7 nm (such as about 6 nm), and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the thickness
of the albumin coating of the nanoparticles under cryo-TEM, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a percentage of albumin monomers among the albumin
on the nanoparticles being about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), a thickness of about 5-7 nm (such
as about 6 nm), and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the thickness of the albumin coating of the nanoparticles under
cryo-TEM, and determining the distribution of rapamycin in a tumor
tissue upon injection of the pharmaceutical composition directly
into the tumor tissue, wherein a percentage of albumin polymer
among the albumin on the nanoparticles being about 15% to about 40%
(such as about 15% to about 20%, about 20% to about 24.5%, about
24.5% to about 30%, about 30% to about 35%, or about 35% to about
40%), a percentage of albumin monomer among the albumin on the
nanoparticles being at least about 40% to about 60% (such as about
40% to about 55%, about 40% to about 54%, about 40% to about 53%,
about 40% to about 52%, about 40% to about 50%, about 40% to about
48%, or about 40% to about 46%), a thickness of about 5-7 nm (such
as about 6 nm), and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the thickness of the albumin
coating of the nanoparticles under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a weight percentage of the albumin in the nanoparticles being about
15% to about 30% (such as about 20% to about 25%, about 15% to
about 24%, or about 15% to about 20%), a thickness of about 5-7 nm
(such as about 6 nm), and an enhanced rapamycin tumor distribution
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
thickness of the albumin coating of the nanoparticles under
cryo-TEM, and determining the distribution of rapamycin in a tumor
tissue upon injection of the pharmaceutical composition directly
into the tumor tissue, wherein a percentage of albumin polymer
among the albumin on the nanoparticles being about 15% to about 40%
(such as about 15% to about 20%, about 20% to about 24.5%, about
24.5% to about 30%, about 30% to about 35%, or about 35% to about
40%), a weight percentage of the albumin in the nanoparticles being
about 15% to about 30% (such as about 20% to about 25%, about 15%
to about 24%, or about 15% to about 20%), a thickness of about 5-7
nm (such as about 6 nm), and an enhanced rapamycin tumor
distribution is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if upon tumor injection it allows rapamycin to spread radially for
a distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
thickness of the albumin coating of the nanoparticles under
cryo-TEM, and determining the distribution of rapamycin in a tumor
tissue upon injection of the pharmaceutical composition directly
into the tumor tissue, wherein a percentage of albumin monomers
among the albumin on the nanoparticles being about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), a weight percentage
of the albumin in the nanoparticles being about 15% to about 30%
(such as about 20% to about 25%, about 15% to about 24%, or about
15% to about 20%), a thickness of about 5-7 nm (such as about 6
nm), and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the thickness of the albumin coating of the
nanoparticles under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), a weight percentage
of the albumin in the nanoparticles being about 15% to about 30%
(such as about 20% to about 25%, about 15% to about 24%, or about
15% to about 20%), a thickness of about 5-7 nm (such as about 6
nm), and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles, determining the thickness of the
albumin coating of the nanoparticles under cryo-TEM, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein an albumin to rapamycin ratio of about 1:2 to about
1:6 in the nanoparticles, a thickness of about 5-7 nm (such as
about 6 nm), and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, determining the
thickness of the albumin coating of the nanoparticles under
cryo-TEM, and determining the distribution of rapamycin in a tumor
tissue upon injection of the pharmaceutical composition directly
into the tumor tissue, wherein a percentage of albumin polymer
among the albumin on the nanoparticles being about 15% to about 40%
(such as about 15% to about 20%, about 20% to about 24.5%, about
24.5% to about 30%, about 30% to about 35%, or about 35% to about
40%), an albumin to rapamycin ratio of about 1:2 to about 1:6 in
the nanoparticles, a thickness of about 5-7 nm (such as about 6
nm), and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, determining the
thickness of the albumin coating of the nanoparticles under
cryo-TEM, and determining the distribution of rapamycin in a tumor
tissue upon injection of the pharmaceutical composition directly
into the tumor tissue, wherein a percentage of albumin monomers
among the albumin on the nanoparticles being about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles, a
thickness of about 5-7 nm (such as about 6 nm), and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight ratio of albumin to rapamycin in the nanoparticles,
determining the thickness of the albumin coating of the
nanoparticles under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as any of about 15% to about 20%, about 20%
to about 24.5%, about 24.5% to about 30%, about 30% to about 35%,
or about 35% to about 40%), and a percentage of albumin monomer
among the albumin on the nanoparticles being at least about 40% to
about 60% (such as about 40% to about 55%, about 40% to about 54%,
about 40% to about 53%, about 40% to about 52%, about 40% to about
50%, about 40% to about 48%, or about 40% to about 46%), an albumin
to rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles,
a thickness of about 5-7 nm (such as about 6 nm), and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the weight ratio of albumin to
rapamycin in the nanoparticles, determining the thickness of the
albumin coating of the nanoparticles under cryo-TEM, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein a weight percentage of the albumin in the
nanoparticles being about 15% to about 30% (such as about 20% to
about 25%, about 15% to about 24%, or about 15% to about 20%), an
albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles, a thickness of about 5-7 nm (such as about 6 nm),
and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles,
determining the thickness of the albumin coating of the
nanoparticles under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%), a weight percentage of the albumin in the
nanoparticles being about 15% to about 30% (such as about 20% to
about 25%, about 15% to about 24%, or about 15% to about 20%), an
albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles, a thickness of about 5-7 nm (such as about 6 nm),
and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles,
determining the thickness of the albumin coating of the
nanoparticles under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin monomers among the albumin on the nanoparticles being about
40% to about 60% (such as about 40% to about 55%, about 40% to
about 54%, about 40% to about 53%, about 40% to about 52%, about
40% to about 50%, about 40% to about 48%, or about 40% to about
46%), a weight percentage of the albumin in the nanoparticles being
about 15% to about 30% (such as about 20% to about 25%, about 15%
to about 24%, or about 15% to about 20%), an albumin to rapamycin
ratio of about 1:2 to about 1:6 in the nanoparticles, a thickness
of about 5-7 nm (such as about 6 nm), and an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the weight ratio of albumin to rapamycin in the
nanoparticles, determining the thickness of the albumin coating of
the nanoparticles under cryo-TEM, and determining the distribution
of rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), a weight percentage
of the albumin in the nanoparticles being about 15% to about 30%
(such as about 20% to about 25%, about 15% to about 24%, or about
15% to about 20%), an albumin to rapamycin ratio of about 1:2 to
about 1:6 in the nanoparticles, a thickness of about 5-7 nm (such
as about 6 nm), and an enhanced rapamycin tumor distribution is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the morphology and thickness of the
albumin coating of the nanoparticles under cryo-TEM, and
determining the distribution of rapamycin in a tumor tissue upon
injection of the pharmaceutical composition directly into the tumor
tissue, wherein an irregular shape of the nanoparticles and a
thickness of about 5-7 nm (such as about 6 nm), and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the morphology
of the nanoparticles and thickness of the albumin coating under
cryo-TEM, and determining the distribution of rapamycin in a tumor
tissue upon injection of the pharmaceutical composition directly
into the tumor tissue, wherein a percentage of albumin polymer
among the albumin on the nanoparticles being about 15% to about 40%
(such as about 15% to about 20%, about 20% to about 24.5%, about
24.5% to about 30%, about 30% to about 35%, or about 35% to about
40%), an irregular shape of the nanoparticles, a thickness of about
5-7 nm (such as about 6 nm), and an enhanced rapamycin tumor
distribution is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if upon tumor injection it allows rapamycin to spread radially for
a distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the morphology
of the nanoparticles and thickness of the albumin coating under
cryo-TEM, and determining the distribution of rapamycin in a tumor
tissue upon injection of the pharmaceutical composition directly
into the tumor tissue, wherein a percentage of albumin monomers
among the albumin on the nanoparticles being about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), an irregular shape of
the nanoparticles, a thickness of about 5-7 nm (such as about 6
nm), and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the morphology of the nanoparticles and thickness of the
nanoparticles under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%), a percentage of albumin monomer among the
albumin on the nanoparticles being at least about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, about 40% to about 52%, about 40% to about 50%, about
40% to about 48%, or about 40% to about 46%), an irregular shape of
the nanoparticles, a thickness of about 5-7 nm (such as about 6
nm), and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the morphology of the
nanoparticles and thickness of the nanoparticles under cryo-TEM,
and determining the distribution of rapamycin in a tumor tissue
upon injection of the pharmaceutical composition directly into the
tumor tissue, wherein a weight percentage of the albumin in the
nanoparticles being about 15% to about 30% (such as about 20% to
about 25%, about 15% to about 24%, or about 15% to about 20%), an
irregular shape of the nanoparticles, a thickness of about 5-7 nm
(such as about 6 nm), and an enhanced rapamycin tumor distribution
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%), a weight percentage of the albumin in the
nanoparticles being about 15% to about 30% (such as about 20% to
about 25%, about 15% to about 24%, or about 15% to about 20%), an
irregular shape of the nanoparticles, a thickness of about 5-7 nm
(such as about 6 nm), and an enhanced rapamycin tumor distribution
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin monomers among the albumin on the nanoparticles being about
40% to about 60% (such as about 40% to about 55%, about 40% to
about 54%, about 40% to about 53%, about 40% to about 52%, about
40% to about 50%, about 40% to about 48%, or about 40% to about
46%), a weight percentage of the albumin in the nanoparticles being
about 15% to about 30% (such as about 20% to about 25%, about 15%
to about 24%, or about 15% to about 20%), an irregular shape of the
nanoparticles, a thickness of about 5-7 nm and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the morphology of the nanoparticles and thickness of
the albumin coating under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), a weight percentage of the albumin in the nanoparticles
being about 15% to about 30% (such as about 20% to about 25%, about
15% to about 24%, or about 15% to about 20%), an irregular shape of
the nanoparticles, a thickness of about 5-7 nm (such as about 6
nm), and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles, determining the morphology of the
nanoparticles and thickness of the albumin coating under cryo-TEM,
and determining the distribution of rapamycin in a tumor tissue
upon injection of the pharmaceutical composition directly into the
tumor tissue, wherein an albumin to rapamycin ratio of about 1:2 to
about 1:6 in the nanoparticles, an irregular shape of the
nanoparticles, a thickness of about 5-7 nm (such as about 6 nm),
and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, determining the
morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%), an albumin to rapamycin ratio of about 1:2
to about 1:6 in the nanoparticles, an irregular shape of the
nanoparticles, a thickness of about 5-7 nm (such as about 6 nm),
and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
ratio of albumin to rapamycin in the nanoparticles, determining the
morphology of the nanoparticles and thickness of the albumin
coating under cryo-TEM, and determining the distribution of
rapamycin in a tumor tissue upon injection of the pharmaceutical
composition directly into the tumor tissue, wherein a percentage of
albumin monomers among the albumin on the nanoparticles being about
40% to about 60% (such as about 40% to about 55%, about 40% to
about 54%, about 40% to about 53%, about 40% to about 52%, about
40% to about 50%, about 40% to about 48%, or about 40% to about
46%), an albumin to rapamycin ratio of about 1:2 to about 1:6 in
the nanoparticles, an irregular shape of the nanoparticles, a
thickness of about 5-7 nm (such as about 6 nm), and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight ratio of albumin to rapamycin in the nanoparticles,
determining the morphology of the nanoparticles and thickness of
the albumin coating under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), an albumin to rapamycin ratio of about 1:2 to about 1:6
in the nanoparticles, an irregular shape of the nanoparticles, a
thickness of about 5-7 nm (such as about 6 nm), and an enhanced
rapamycin tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the weight ratio of albumin to
rapamycin in the nanoparticles, determining the morphology of the
nanoparticles and thickness of the albumin coating under cryo-TEM,
and determining the distribution of rapamycin in a tumor tissue
upon injection of the pharmaceutical composition directly into the
tumor tissue, wherein a weight percentage of the albumin in the
nanoparticles being about 15% to about 30% (such as about 20% to
about 25%, about 15% to about 24%, or about 15% to about 20%), an
albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles, an irregular shape of the nanoparticles, a thickness
of about 5-7 nm (such as about 6 nm), and an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
polymers among the albumin on the nanoparticles. In some
embodiments, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition. In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light
scattering). In some embodiments, the method further comprises
determining the polydispersity index of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin polymers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles,
determining the morphology of the nanoparticles and thickness of
the albumin coating under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a weight
percentage of the albumin in the nanoparticles being about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%), an albumin to rapamycin ratio of about
1:2 to about 1:6 in the nanoparticles, an irregular shape of the
nanoparticles, a thickness of about 5-7 nm (such as about 6 nm),
and an enhanced rapamycin tumor distribution is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radially for a distance that is greater
than (for example more than about any of 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example injected
at the rapamycin amount of about 12 .mu.g (such as at about 4
mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In some
embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin monomers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin dimers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin oligomers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
among the albumin on the nanoparticles, determining the weight
percentage of the albumin in the nanoparticles, determining the
weight ratio of albumin to rapamycin in the nanoparticles,
determining the morphology of the nanoparticles and thickness of
the albumin coating under cryo-TEM, and determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor tissue, wherein
a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%), a weight percentage of the albumin in
the nanoparticles being about 15% to about 30% (such as about 20%
to about 25%, about 15% to about 24%, or about 15% to about 20%),
an albumin to rapamycin ratio of about 1:2 to about 1:6 in the
nanoparticles, an irregular shape of the nanoparticles, a thickness
of about 5-7 nm (such as about 6 nm), and an enhanced rapamycin
tumor distribution is indicative of suitability of the
pharmaceutical composition for medical use. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if upon tumor injection it allows rapamycin to
spread radially for a distance that is greater than (for example
more than about any of 1.1.times., 1.2.times., 1.3.times.,
1.4.times., 1.5.times., 1.6.times., 1.7.times., 1.8.times.,
2.times., 3.times., 4.times., 5.times., 6.times., 7.times.,
8.times., or more of) that of a solvent-based rapamycin (such as
rapamycin in DMSO) formulation under the same assay conditions. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows rapamycin to spread radically for more than about 700 .mu.m
(such as more than about any of 700 .mu.m, 800 .mu.m, 900 .mu.m,
1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within about 24 hours after
the composition (such as a pharmaceutical composition) is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor). In some embodiments, the method further
comprises determining the solubility of the pharmaceutical
composition (including determining solubility after storage). In
some embodiments, a percentage of albumin monomer among the albumin
on the nanoparticles being less than about 51% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers and oligomers
among the albumin on the nanoparticles being more than about 35% is
indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, a percentage of albumin polymers
among the albumin on the nanoparticles being more than about 17%
and a percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight percentage of the albumin in the nanoparticles,
determining the weight ratio of albumin to rapamycin in the
nanoparticles, determining the morphology of the nanoparticles and
thickness of the albumin coating under cryo-TEM, and determining
the distribution of rapamycin in a tumor tissue upon injection of
the pharmaceutical composition directly into the tumor tissue,
wherein a percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%), a percentage of
albumin monomer among the albumin on the nanoparticles being at
least about 40% to about 60% (such as about 40% to about 55%, about
40% to about 54%, about 40% to about 53%, about 40% to about 52%,
about 40% to about 50%, about 40% to about 48%, or about 40% to
about 46%), a weight percentage of the albumin in the nanoparticles
being about 15% to about 30% (such as about 20% to about 25%, about
15% to about 24%, or about 15% to about 20%), an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles, an
irregular shape of the nanoparticles, a thickness of about 5-7 nm
(such as about 6 nm), and an enhanced rapamycin tumor distribution
is indicative of suitability of the pharmaceutical composition for
medical use. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if upon
tumor injection it allows rapamycin to spread radially for a
distance that is greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin (such as rapamycin in DMSO) formulation
under the same assay conditions. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if upon tumor injection it allows rapamycin to spread
radically for more than about 700 .mu.m (such as more than about
any of 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or
1200 .mu.m) within about 24 hours after the composition (such as a
pharmaceutical composition) is injected into a tumor tissue (for
example injected at the rapamycin amount of about 12 .mu.g (such as
at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In
some embodiments, the method further comprises determining the
solubility of the pharmaceutical composition (including determining
solubility after storage). In some embodiments, a percentage of
albumin monomer among the albumin on the nanoparticles being less
than about 51% is indicative of suitability of the pharmaceutical
composition for medical use. In some embodiments, a percentage of
albumin polymers and oligomers among the albumin on the
nanoparticles being more than about 35% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 17% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 54% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a percentage of albumin polymers among the
albumin on the nanoparticles being more than about 18% and a
percentage of albumin monomer among the albumin on the
nanoparticles being less than about 55% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, a ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers being more than about 65% is indicative of
suitability of the pharmaceutical composition for medical use. In
some embodiments, the method further comprises determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage). In some
embodiments, the method further comprises determining the rapamycin
recovery following a 0.2 micron filtration (including determining
recovery after storage). In some embodiments, the method further
comprises determining binding affinity of albumin to rapamycin in
the composition (such as a pharmaceutical composition) (for example
by equilibrium dialysis, FTIR, NMR, or a combination thereof). In
some embodiments, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin monomers,
dimers, oligomers, or polymers among the total albumin in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the particle size of the nanoparticles (for
example by dynamic light scattering). In some embodiments, the
method further comprises determining the polydispersity index of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the span of
size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the surface
potential of the nanoparticles. In some embodiments, the method
further comprises determining the percentage of the rapamycin in
the nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC). In some
embodiments, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition (for
example by size-exclusion chromatography). In some embodiments, the
method further comprises determining the in vitro release kinetics
of the composition (such as a pharmaceutical composition). In some
embodiments, the method further comprises determining the stability
of the pharmaceutical composition (including determining stability
after storage).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the solubility, rapamycin
crystallinity, and rapamycin recovery following a 0.2 micron
filtration of the pharmaceutical composition, wherein a solubility
of about 50 .mu.g/ml to about 100 .mu.g/ml in a 5% human albumin
solution, a non-crystalline state, and a recovery of at least about
80% is indicative of suitability of the pharmaceutical composition
for medical use. In some embodiments, the solubility, rapamycin
crystallinity, and/or rapamycin recovery are determined after
storage (for example after storage for at least about 6 hours, 12
hours, 18 hours, 24 hours, 36 hours, 48 hours, or 72 hours, such as
at room temperature, under refrigerated condition, or at 40.degree.
C.). In some embodiments, the rapamycin crystallinity is determined
by X-ray diffraction and/or polarized light microscopy. In some
embodiments, the method further comprises determining binding
affinity of albumin to rapamycin in the composition (such as a
pharmaceutical composition) (for example by equilibrium dialysis,
FTIR, NMR, or a combination thereof). In some embodiments, the
method further comprises determining the surface-to-volume ratio of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the method further comprises determining the
percentage of albumin monomers among the albumin on the
nanoparticles. In some embodiments, the method further comprises
determining the percentage of albumin dimers among the albumin on
the nanoparticles. In some embodiments, the method further
comprises determining the percentage of albumin oligomers among the
albumin on the nanoparticles. In some embodiments, the method
further comprises determining the percentage of albumin polymers
among the albumin on the nanoparticles. In some embodiments, the
method further comprises determining the percentage of albumin
monomers, dimers, oligomers, or polymers among the total albumin in
the pharmaceutical composition. In some embodiments, the method
further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering). In some
embodiments, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition. In some embodiments, the method further comprises
determining the span of size distribution
((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the nanoparticles in the
pharmaceutical composition. In some embodiments, the method further
comprises determining the surface potential of the nanoparticles.
In some embodiments, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC). In some embodiments, the method further
comprises determining the percentage of the albumin that is in the
non-nanoparticle portion among the total albumin in the
pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining the in vitro release kinetics of the composition (such
as a pharmaceutical composition). In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
The different determination steps described above may be carried
out in various combinations in a given method.
For example, in some embodiments, there is provided a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
monomers and polymers among the albumin on the nanoparticles,
determining the weight percentage of the albumin in the
nanoparticles, determining the solubility of the pharmaceutical
composition (including determining solubility after storage),
determining the rapamycin crystallinity of the pharmaceutical
composition (for example by X-ray diffraction and/or polarized
light microscopy, including determining crystallinity after
storage), and determining the rapamycin recovery following a 0.2
micron filtration (including determining recovery after storage).
In some embodiments, the method further comprises determining
binding affinity of albumin to rapamycin in the composition (such
as a pharmaceutical composition) (for example by equilibrium
dialysis, FTIR, NMR, or a combination thereof). In some
embodiments, the method further comprises determining the size of
the nanoparticles (for example by dynamic light scattering) and/or
size distribution of the nanoparticles. In some embodiments, the
method further comprises determining the stability of the
pharmaceutical composition (including determining stability after
storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers
and polymers among the albumin on the nanoparticles, determining
the weight ratio of albumin to rapamycin in the nanoparticles,
determining the morphology of the nanoparticles and thickness of
the albumin coating under cryo-TEM, determining the solubility of
the pharmaceutical composition (including determining solubility
after storage), determining the rapamycin crystallinity of the
pharmaceutical composition (for example by X-ray diffraction and/or
polarized light microscopy, including determining crystallinity
after storage), and determining the rapamycin recovery following a
0.2 micron filtration (including determining recovery after
storage). In some embodiments, the method further comprises
determining binding affinity of albumin to rapamycin in the
composition (such as a pharmaceutical composition) (for example by
equilibrium dialysis, FTIR, NMR, or a combination thereof). In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light scattering)
and/or size distribution of the nanoparticles. In some embodiments,
the method further comprises determining the stability of the
pharmaceutical composition (including determining stability after
storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the weight percentage of the albumin
in the nanoparticles, determining the morphology of the
nanoparticles and thickness of the albumin coating under cryo-TEM,
determining the solubility of the pharmaceutical composition
(including determining solubility after storage), determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage), and determining
the rapamycin recovery following a 0.2 micron filtration (including
determining recovery after storage). In some embodiments, the
method further comprises determining binding affinity of albumin to
rapamycin in the composition (such as a pharmaceutical composition)
(for example by equilibrium dialysis, FTIR, NMR, or a combination
thereof). In some embodiments, the method further comprises
determining the size of the nanoparticles (for example by dynamic
light scattering) and/or size distribution of the nanoparticles. In
some embodiments, the method further comprises determining the
stability of the pharmaceutical composition (including determining
stability after storage). In some embodiments, the method further
comprises determining tumor distribution of rapamycin upon
administration in vivo (for example by determining tumor
distribution of rapamycin upon injection of the pharmaceutical
composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the percentage of albumin monomers,
dimers, oligomers, and polymers among the albumin on the
nanoparticles, determining the weight percentage of the albumin in
the nanoparticles, determining the morphology of the nanoparticles
and thickness of the albumin coating under cryo-TEM, determining
the surface-to-volume ratio of the nanoparticles in the
pharmaceutical composition, and determining the percentage of
albumin monomers, dimers, oligomers, or polymers among the total
albumin in the pharmaceutical composition. In some embodiments, the
method further comprises determining binding affinity of albumin to
rapamycin in the composition (such as a pharmaceutical composition)
(for example by equilibrium dialysis, FTIR, NMR, or a combination
thereof). In some embodiments, the method further comprises
determining the particle size of the nanoparticles (for example by
dynamic light scattering) and/or size distribution of the
nanoparticles. In some embodiments, the method further comprises
determining the stability of the pharmaceutical composition
(including determining stability after storage). In some
embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
monomers, dimers, oligomers, and polymers among the albumin on the
nanoparticles, determining the weight percentage of the albumin in
the nanoparticles, determining the rapamycin crystallinity of the
pharmaceutical composition (for example by X-ray diffraction and/or
polarized light microscopy, including determining crystallinity
after storage), determining the rapamycin recovery following a 0.2
micron filtration (including determining recovery after storage),
determining the percentage of albumin monomers, dimers, oligomers,
or polymers among the total albumin in the pharmaceutical
composition, determining the percentage of the rapamycin in the
nanoparticles among the total rapamycin in the pharmaceutical
composition (for example by reversed-phase HPLC), and determining
the percentage of the albumin that is in the non-nanoparticle
portion among the total albumin in the pharmaceutical composition
(for example by size-exclusion chromatography). In some
embodiments, the method further comprises determining binding
affinity of albumin to rapamycin in the composition (such as a
pharmaceutical composition) (for example by equilibrium dialysis,
FTIR, NMR, or a combination thereof). In some embodiments, the
method further comprises determining the particle size of the
nanoparticles (for example by dynamic light scattering) and/or size
distribution of the nanoparticles. In some embodiments, the method
further comprises determining the stability of the pharmaceutical
composition (including determining stability after storage). In
some embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
monomers, dimers, oligomers, and polymers among the albumin on the
nanoparticles, determining the weight percentage of the albumin in
the nanoparticles, determining the weight ratio of albumin to
rapamycin in the nanoparticles, determining the morphology of the
nanoparticles and thickness of the albumin coating under cryo-TEM,
determining the solubility of the pharmaceutical composition
(including determining solubility after storage), determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage), determining the
rapamycin recovery following a 0.2 micron filtration (including
determining recovery after storage), determining the percentage of
albumin monomers, dimers, oligomers, or polymers among the total
albumin in the pharmaceutical composition, determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC), and determining the percentage of the albumin
that is in the non-nanoparticle portion among the total albumin in
the pharmaceutical composition (for example by size-exclusion
chromatography). In some embodiments, the method further comprises
determining binding affinity of albumin to rapamycin in the
composition (such as a pharmaceutical composition) (for example by
equilibrium dialysis, FTIR, NMR, or a combination thereof). In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light scattering)
and/or size distribution of the nanoparticles. In some embodiments,
the method further comprises determining the stability of the
pharmaceutical composition (including determining stability after
storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
monomers, dimers, oligomers, and polymers among the albumin on the
nanoparticles, determining the weight percentage of the albumin in
the nanoparticles, determining the weight ratio of albumin to
rapamycin in the nanoparticles, determining the morphology of the
nanoparticles and thickness of the albumin coating under cryo-TEM,
determining the solubility of the pharmaceutical composition
(including determining solubility after storage), determining the
rapamycin crystallinity of the pharmaceutical composition (for
example by X-ray diffraction and/or polarized light microscopy,
including determining crystallinity after storage), determining the
rapamycin recovery following a 0.2 micron filtration (including
determining recovery after storage), determining the percentage of
albumin monomers, dimers, oligomers, or polymers among the total
albumin in the pharmaceutical composition, determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition (for example by
reversed-phase HPLC), determining the percentage of the albumin
that is in the non-nanoparticle portion among the total albumin in
the pharmaceutical composition (for example by size-exclusion
chromatography), determining the particle size of the nanoparticles
(for example by dynamic light scattering) and/or size distribution
of the nanoparticles, and determining the stability of the
pharmaceutical composition (including determining stability after
storage). In some embodiments, the method further comprises
determining binding affinity of albumin to rapamycin in the
composition (such as a pharmaceutical composition) (for example by
equilibrium dialysis, FTIR, NMR, or a combination thereof). In some
embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the solubility, rapamycin
crystallinity, and rapamycin recovery following a 0.2 micron
filtration of the pharmaceutical composition, determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition,
determining the percentage of the rapamycin in the nanoparticles
among the total rapamycin in the pharmaceutical composition (for
example by reversed-phase HPLC), and determining the percentage of
the albumin that is in the non-nanoparticle portion among the total
albumin in the pharmaceutical composition (for example by
size-exclusion chromatography). In some embodiments, the
solubility, rapamycin crystallinity, and/or rapamycin recovery are
determined after storage (for example after storage for at least
about 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 hours, or
72 hours, such as at room temperature, under refrigerated
condition, or at 40.degree. C.). In some embodiments, the rapamycin
crystallinity is determined by X-ray diffraction and/or polarized
light microscopy. In some embodiments, the method further comprises
determining binding affinity of albumin to rapamycin in the
composition (such as a pharmaceutical composition) (for example by
equilibrium dialysis, FTIR, NMR, or a combination thereof). In some
embodiments, the method further comprises determining the particle
size of the nanoparticles (for example by dynamic light scattering)
and/or size distribution of the nanoparticles. In some embodiments,
the method further comprises determining the stability of the
pharmaceutical composition (including determining stability after
storage). In some embodiments, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo (for example by determining tumor distribution of rapamycin
upon injection of the pharmaceutical composition directly into the
tumor tissue).
In some embodiments, there is provided a method of assessing
suitability of a pharmaceutical composition for medical use in a
human individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: determining the solubility, rapamycin
crystallinity, and rapamycin recovery following a 0.2 micron
filtration of the pharmaceutical composition, determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition,
determining the percentage of the rapamycin in the nanoparticles
among the total rapamycin in the pharmaceutical composition (for
example by reversed-phase HPLC), determining the percentage of the
albumin that is in the non-nanoparticle portion among the total
albumin in the pharmaceutical composition (for example by
size-exclusion chromatography), determining the particle size of
the nanoparticles (for example by dynamic light scattering) and/or
size distribution of the nanoparticles, and determining the
stability of the pharmaceutical composition (including determining
stability after storage). In some embodiments, the solubility,
rapamycin crystallinity, and/or rapamycin recovery are determined
after storage (for example after storage for at least about any of
6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 hours, or 72
hours, such as at room temperature, under refrigerated condition,
or at about 40.degree. C.). In some embodiments, the rapamycin
crystallinity is determined by X-ray diffraction and/or polarized
light microscopy. In some embodiments, the method further comprises
determining binding affinity of albumin to rapamycin in the
composition (such as a pharmaceutical composition) (for example by
equilibrium dialysis, FTIR, NMR, or a combination thereof). In some
embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
Determination of Albumin Oligomeric Status on Nanoparticles
The methods of the present application in some embodiments require
determination of oligomeric status (e.g., polymers, monomers,
dimers, and/or oligomers) of the albumin on the nanoparticles. The
oligomeric status of the albumin on the nanoparticles may impact
the particle stability, solubility, dissolution rate, and in vivo
distribution. Further, because albumin-rapamycin binding is greater
for crosslinked albumin (namely, albumin polymers, oligomers, and
dimers) than albumin monomers, the oligomeric status of the albumin
on the nanoparticles may also affect in vivo behavior of the
albumin-based rapamycin nanoparticle composition.
In some embodiments, the oligomeric status of the albumin is
determined by size-exclusion chromatography, such as gel permeation
chromatography or HPLC size-exclusion methods, or polyacrylamide
gel electrophoresis (such as sodium dodecyl sulfate polyacrylamide
gel electrophoresis, SDS-PAGE). In some embodiments, the oligomeric
status is determined by isolating the albumin on the nanoparticles
in the pharmaceutical composition by, for example,
ultracentrifugation or gel filtration chromatography, and further
analyzing the albumin on the nanoparticles by, for example,
size-exclusion chromatography. The different classes of albumins
can be determined based on differing retention times of albumins
when subject to a chromatography (such as size-exclusion
chromatography, e.g., gel permeation chromatography). In some
embodiments, the different classes of albumins can be determined
based on RRT. In some embodiments, the oligomeric status is
determined upon reconstitution of the pharmaceutical composition.
In some embodiments, the oligomeric status is determined upon
storage of the pharmaceutical composition.
In some embodiments, the size-exclusion chromatography method used
is capable of separating monomeric albumin from dimeric albumin,
oligomeric albumin, and polymeric albumin. In some embodiments, the
size-exclusion chromatography method used is capable of separating
dimeric albumin from monomeric albumin, oligomeric albumin, and
polymeric albumin. In some embodiments, the size-exclusion
chromatography method used is capable of separating oligomeric
albumin from monomeric albumin, dimeric albumin, and polymeric
albumin. In some embodiments, the size-exclusion chromatography
method used is capable of separating polymeric albumin from
monomeric albumin, dimeric albumin, and polymeric albumin. In some
embodiments, the size-exclusion chromatography method used is
capable of separating all four categories of albumin on the
nanoparticles (e.g., monomeric, dimeric, oligomeric,
polymeric).
In some embodiments, when determining the oligomeric status of the
albumin, the separation range for the size-exclusion chromatography
is about 10,000 daltons to about 500,000 daltons. In some
embodiments, the size-exclusion chromatography is run with a TSKgel
G3000 SWXL column. In some embodiments, the size-exclusion
chromatography is run with a column of TOSOH TSKgel G3000 SWXL,
7.8.times.300 mm, 5 .mu.m or equivalent. In some embodiments, the
size-exclusion chromatography is run with a flow rate of about 1
mL/min. In some embodiments, the size-exclusion chromatography is
run at ambient temperature. In some embodiments, the size-exclusion
chromatography is run with a column of TOSOH TSKgel G3000 SWXL,
7.8.times.300 mm, 5 .mu.m or equivalent, at a flow rate of about 1
mL/min at room temperature.
The percentage of the albumin on the nanoparticles that is in the
form of a monomer, polymer, dimer, and/or oligomer can be
determined by comparing the amount of monomeric, polymeric,
dimeric, and/or oligomeric albumin on the nanoparticles with the
total amount of the albumin on the nanoparticles.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the percentage of
monomeric albumin among the albumin on the nanoparticles is about
40% to about 60%. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
percentage of monomeric albumin among the albumin on the
nanoparticles is about any one of 30%, 35%, 40%, 45%, 50%, 55%,
60%, 65%, or 70%. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
percentage of monomeric albumin among the albumin on the
nanoparticles is about any one of 20-30%, 30-40%, 40-50%, 50-60%,
60-70%, 70-80%, 20-40%, 40-60%, 60-80%, 20-50%, 50-80%, 35-40%,
40-45%, 45-50%, 50-55%, 55-60%, 60-65%, 35-45%, 45-55%, 55-65%,
40-55%, or 45-60%.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the percentage of
dimeric albumin among the albumin on the nanoparticles is about 15%
to about 30% (such as about 20% to about 25%, about 15% to about
24%, or about 15% to about 20%). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the percentage of dimeric albumin among the albumin
on the nanoparticles is about any one of 10%, 11%, 12%, 13%, 14%,
15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, or 30%. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the percentage of
dimeric albumin among the albumin on the nanoparticles is about any
of 10-12%, 12-14%, 14-15%, 15-16%, 16-17%, 17-18%, 18-19%, 19-20%,
20-21%, 21-23%, 23-25%0, 10-15%, 15-20%, 20-25%, 15-17%, 17-19%,
15-15.5%, 15.5-16%, 16-16.5%, 16.5-17%, 17-17.5%, 17.5-18%,
18-18.5%, 18.5-19%, 19-19.5%, 19.5-20%, 15.5-16.5%, 16.5-17.5%,
17.5-18.5%, 18.5-19.5%, 15-16.5%, 16-17.5%, 17-18.5%, 18-19.5%,
16.5-19%, or 17.5-20%.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the percentage of
oligomeric albumin among the albumin on the nanoparticles is about
7% to about 15%. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
percentage of oligomeric albumin among the albumin on the
nanoparticles is about any one of 5%, 6%, 7%, 8%, 9%, 10%, 11%,
12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or 25%. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the percentage of oligomeric albumin
among the albumin on the nanoparticles is about any one of about
any one of 5-6%, 6-7%, 7-8%, 8-9%, 9-10%, 10-11%, 11-12%, 12-13%,
13-14%, 14-15%, 15-16%, 16-17%, 17-20%, 20-25%, 5-7%, 7-9%, 9-11%,
11-13%, 13-15%, 7-10%, 10-13%, 7-12%, 12-15%, 10-15%, or
15-20%.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the percentage of
polymeric albumin among the albumin on the nanoparticles is about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%). In some embodiments, the composition (such
as a pharmaceutical composition) is suitable for medical use if the
percentage of polymeric albumin among the albumin on the
nanoparticles is about any one of 15%, 16%, 17%, 18%, 19%, 20%,
22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, or 40%. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the percentage of polymeric albumin
among the albumin on the nanoparticles is any of about 15% to about
16%, about 16% to about 17%, about 17% to about 18%, about 18% to
about 19%, about 19% to about 20%, about 20% to about 21%, about
21% to about 22%, about 22% to about 23%, about 23% to about 24%,
about 24% to about 25%, about 25% to about 26%, about 26% to about
27%, about 27% to about 28%, about 28% to about 29%, about 29% to
about 30%, about 30% to about 35%, about 35% to about 40%, about
15% to about 18%, about 18% to about 20%, about 20% to about 23%,
about 23% to about 25%, about 25% to about 30%, about 30% to about
40%, about 15% to about 20%, about 20% to about 24.5%, about 24.5%
to about 30%, about 15% to about 24.5%, about 15% to about 18.5%,
or about 15% to about 40% (such as about 15% to about 20%, about
20% to about 24.5%, about 24.5% to about 30%, about 30% to about
35%, or about 35% to about 40%). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the percentage of polymeric albumin among the
albumin on the nanoparticles is about 23.6-24.7%, or more than
about 29.7%. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
percentage of polymeric albumin among the albumin on the
nanoparticles is about 23.6-24.7%. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the percentage of oligomeric albumin among the
albumin on the nanoparticles is about any one of 10-20%, 20-30%,
30-40%, 15-25%, 25-35%, 35-40%, 21-22%, 22-23%, 23-24%, 24-25%,
25-26%, 26-28%, 28-30%, 20-23%, 23-25%, 25-30%, 23-23.2%,
23.2-23.4%, 23.4-23.6%, 23.6-23.8%, 23.8-24%, 24-24.2%, 24.2-24.4%,
24.4-24.6%, 24.6-24.8%, 24.8-25%, 23-23.4%, 23.4-23.8%, 23.8-24.2%,
24.2-24.6%, 24.6%-25%, 23-23.5%, 23.5-24%, 24-24.5%, 24.5-25%,
23-23.6%, 23.6-24.2%, 24.2-24.7%, or 23.6-24.7%.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the percentage of
polymeric albumin among the albumin on the nanoparticles is more
than about 29.7%. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
percentage of polymeric albumin among the albumin on the
nanoparticles is about 30% to about 32%, about 32% to about 34%,
about 34% to about 36%, about 36% to about 38%, or about 38% to
about 40%.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if about 15% to about 40%
(such as about 15% to about 20%, about 20% to about 24.5%, about
24.5% to about 30%, about 30% to about 35%, or about 35% to about
40%) of the albumin on the nanoparticles is in the form of
polymers, and about 40% to about 60% of the albumin on the
nanoparticles is in the form of monomers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if at least about any one of 10%, 12%, 13%, 14%, 15%,
16%, 17%, 18%, 19%, 20%, 22%, 23%, 24%, 25%, of the albumin on the
nanoparticles is in the form of polymers, and about any one of 30%,
35%, 40%, 45%, 50%, 55%, 60%, or 65% of the albumin on the
nanoparticles is in the form of monomers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about any one of 10-20%, 20-30%, 30-40%, 15-25%,
25-35%, 35-40%, 21-22%, 22-23%, 23-24%, 24-25%, 25-26%, 26-28%,
28-30%, 20-23%, 23-25%, 25-30%, 23-23.2%, 23.2-23.4%, 23.4-23.6%,
23.6-23.8%, 23.8-24%, 24-24.2%, 24.2-24.4%, 24.4-24.6%, 24.6-24.8%,
24.8-25%, 23-23.4%, 23.4-23.8%, 23.8-24.2%, 24.2-24.6%, 24.6%-25%,
23-23.5%, 23.5-24%, 24-24.5%, 24.5-25%, 23-23.6%, 23.6-24.2%,
24.2-24.7%, or 23.6-24.7% of the albumin on the nanoparticles is in
the form of polymers, and about any one of 20-30%, 30-40%, 40-50%,
50-60%, 60-70%, 70-80%, 20-40%, 40-60%, 60-80%, 20-50%, 50-80%,
35-40%, 40-45%, 45-50%, 50-55%, 55-60%, 60-65%, 35-45%, 45-55%,
55-65%, 40-55%, or 45-60% of the albumin on the nanoparticles is in
the form of monomers. In some embodiments, the composition (such as
a pharmaceutical composition) is suitable for medical use if about
30% to about 32%, about 32% to about 34%, about 34% to about 36%,
about 36% to about 38%, or about 38% to about 40% of the albumin on
the nanoparticles is in the form of polymers, and about any one of
20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 20-40%, 40-60%,
60-80%, 20-50%, 50-80%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%,
60-65%, 35-45%, 45-55%, 55-65%, 40-55%, or 45-60% of the albumin on
the nanoparticles is in the form of monomers.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if about 40% to about 60%
(such as about 40% to about 55%, about 40% to about 54%, about 40%
to about 53%, or about 40% to about 52%, about 40% to about 50%,
about 40% to about 48%, or about 40% to about 46%) of the albumin
on the nanoparticles is in the form of monomers. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if about any one of 30%, 35%, 40%, 45%,
50%, 55%, 60%, 65%, or 70% of the albumin on the nanoparticles is
in the form of monomers. In some embodiments, the composition (such
as a pharmaceutical composition) is suitable for medical use if
about any one of 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%,
20-40%, 40-60%, 60-80%, 20-50%, 50-80%, 35-40%, 40-45%, 45-50%,
50-55%, 55-60%, 60-65%, 35-45%, 45-55%, 55-65%, 40-55%, or 45-60%
of the albumin on the nanoparticles is in the form of monomers. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if less than about 52% of
the albumin on the nanoparticles are in the form of monomers.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if about 7% to about 15%
of the albumin on the nanoparticles is in the form of oligomers. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if about any one of 5%,
6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%,
20%, or 25% of the albumin on the nanoparticles is in the form of
oligomers. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if about
any one of 5-6%, 6-7%, 7-8%, 8-9%, 9-10%, 10-11%, 11-12%, 12-13%,
13-14%, 14-15%, 15-16%, 16-17%, 17-20%, 20-25%, 5-7%, 7-9%, 9-11%,
11-13%, 13-15%, 7-10%, 10-13%, 7-12%, 12-15%, 10-15%, or 15-20% of
the albumin on the nanoparticles is in the form of oligomers.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if about 15% to about 40%
(such as any of about 15% to about 20%, about 20% to about 24.5%,
about 24.5% to about 30%, about 30% to about 35%, or about 35% to
about 40%) of the albumin on the nanoparticles is in the form of
polymers, and about 40% to about 60% of the albumin on the
nanoparticles is in the form of monomers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about any one of 15%, 16%, 17%, 18%, 19%, 20%, 22%,
23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, or 40% of the albumin
on the nanoparticles is in the form of polymers, and about any one
of 30%, 35%, 40%, 45%, 50%, 55%, 60%, or 65% of the albumin on the
nanoparticles is in the form of monomers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about any one of 10-20%, 20-30%, 30-40%, 15-25%,
25-35%, 35-40%, 21-22%, 22-23%, 23-24%, 24-25%, 25-26%, 26-28%,
28-30%, 20-23%, 23-25%, 25-30%, 23-23.2%, 23.2-23.4%, 23.4-23.6%,
23.6-23.8%, 23.8-24%, 24-24.2%, 24.2-24.4%, 24.4-24.6%, 24.6-24.8%,
24.8-25%, 23-23.4%, 23.4-23.8%, 23.8-24.2%, 24.2-24.6%, 24.6%-25%,
23-23.5%, 23.5-24%, 24-24.5%, 24.5-25%, 23-23.6%, 23.6-24.2%,
24.2-24.7%, or 23.6-24.7% of the albumin on the nanoparticles is in
the form of polymers, and about any one of 20-30%, 30-40%, 40-50%,
50-60%, 60-70%, 70-80%, 20-40%, 40-60%, 60-80%, 20-50%, 50-80%,
35-40%, 40-45%, 45-50%, 50-55%, 55-60%, 60-65%, 35-45%, 45-55%,
55-65%, 40-55%, or 45-60% of the albumin on the nanoparticles is in
the form of monomers. In some embodiments, the composition (such as
a pharmaceutical composition) is suitable for medical use if about
30% to about 32%, about 32% to about 34%, about 34% to about 36%,
about 36% to about 38%, or about 38% to about 40% of the albumin on
the nanoparticles is in the form of polymers, and about any one of
20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 20-40%, 40-60%,
60-80%, 20-50%, 50-80%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%,
60-65%, 35-45%, 45-55%, 55-65%, 40-55%, or 45-60% of the albumin on
the nanoparticles is in the form of monomers. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if more than about 35% of albumin on the
nanoparticles are in the forms of polymers and oligomers. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if less than about 54% monomers. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if more than about 35% of
albumin on the nanoparticles are in the forms of polymers and
oligomers. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if less
than about 54% of the albumin on the nanoparticles are in the form
of monomers, and more than about 11% of the albumin in the
nanoparticles are in the form of polymers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if less than about 55% of the albumin on the
nanoparticles are in the form of monomers, and more than about 18%
of the albumin in the nanoparticles are in the form of polymers. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the ratio of albumin on
the nanoparticles in the forms of polymers and oligomers to the
albumin on the nanoparticles in the form of monomers is more than
about 62%.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if less than about 51%
including for example less than about any of 51%, 50%, 49%, 48%,
47%, 46%, 45%, 44%, 43%, or 42% albumin on the nanoparticles are in
the form of monomers. In some embodiments, the composition (such as
a pharmaceutical composition) is suitable for medical use if more
than about 30% including for example more than about any of 30%,
31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% of the albumin
on the nanoparticles are in the forms of polymers and oligomers. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if less than about 52% of
the albumin on the nanoparticles are in the form of monomers, and
more than about 30% (such as more than about 35%) of the albumin on
the nanoparticles are in the form of polymers and oligomers. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if less than about 54% of
the albumin on the nanoparticles are in the form of monomers, and
more than about 17% of the albumin in the nanoparticles are in the
form of polymers. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if less
than about 55% of the albumin on the nanoparticles are in the form
of monomers, and more than about 18% of the albumin in the
nanoparticles are in the form of polymers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the ratio of albumin on the nanoparticles in the
forms of polymers and oligomers to the albumin on the nanoparticles
in the form of monomers is more than about 65% including for
example more than about any of 65%, 66%, 67%, 68%, 69%, 70%, 71%,
72%, 73%, 74%, 75%, or 76%.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the percentage of
albumin on the nanoparticles in the form of monomers minus the
percentage of albumin on the nanoparticles in the form of dimers is
less than about 30% including for example less than about any of
30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40, 41%, 42%,
43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50%. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the percentage of albumin on the nanoparticles in
the form of monomers plus the percentage of albumin on the
nanoparticles in the form of oligomers is less than about 56% to
about 58%. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
percentage of albumin on the nanoparticles in the form of monomers
plus the percentage of albumin on the nanoparticles in the form of
oligomers is less than about any of 56%, 57, 58, 59%, 60%. 61%.
62%. 63%, 64%, or 65%. In some embodiments, the composition (such
as a pharmaceutical composition) is suitable for medical use if the
percentage of albumin on the nanoparticles in the form of monomers
minus the percentage of albumin on the nanoparticles in the form of
polymers is less than about 20% to about 22%. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if the percentage of albumin on the nanoparticles
in the form of monomers minus the percentage of albumin on the
nanoparticles in the form of polymers is less than about any of
20%, 21%, 22%, 23%, 24%, 25%, 26%, 27% or 28%. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if the percentage of albumin on the nanoparticles
in the form of dimers plus the percentage of albumin on the
nanoparticles in the form of polymers is greater than about 42%
including for example greater than about any of 42%, 43%, 44%, 45%,
46%, 47%, 48%, 49%, or 50%. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if the ratio of albumin on the nanoparticles in the forms of
polymers and oligomers to the albumin on the nanoparticles in the
form of monomers minus dimers is more than about 88%. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the ratio of albumin on the
nanoparticles in the forms of polymers and oligomers to the albumin
on the nanoparticles in the form of monomers minus dimers is more
than about any of 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,
84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,
97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 05%, 106%, 107%, 108%,
109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%,
or 120%.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the weight ratio of
albumin on the nanoparticles in the forms of polymers and oligomers
to the albumin on the nanoparticles in the form of monomers is more
than about 6:10. In some embodiments, the weight ratio of albumin
on the nanoparticles in the forms of polymers and oligomers to the
albumin on the nanoparticles in the form of monomers is more than
about any of 6:10, 6.2:10, 6.4:10; 6.6:10, 6.8:10, 7.0:10, 7.2:10;
7.4:10; 7.6:10; or 7.8:10. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if the weight ratio of albumin dimers to albumin monomers on the
nanoparticles is more than about any of 1.6:10, 2.0:10, 2.5:10,
3.0:10, or 3.5:10. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
weight ratio of albumin oligomers to albumin monomers on the
nanoparticles is more than about any of 0.5:10; 1:10, 1.2:10,
1.4:10, or 1.6:10. In some embodiments, the weight ratio of albumin
polymers to albumin monomers on the nanoparticles is less than
about any of 5.8:10, or 5.7:10. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the weight ratio of albumin polymers to albumin
dimers on the nanoparticles is less than about any of 40:10, 30:10,
or 20:10. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
weight ratio of albumin oligomers to albumin dimers on the
nanoparticles is more than about any of 3.3:10, 3.5:10, 3.6:10,
3.8:10, 4.0:10, 4.2:10, or 4.4:10. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the weight ratio of albumin polymers to albumin
oligomers on the nanoparticles is less than about any of 120:10,
100:10, 80:10, 60:10, or 40:10.
Determination of Weight Percentage of Albumin in the
Nanoparticles
The methods of the present application in some embodiments require
determination of the weight percentage of the albumin in the
nanoparticles.
Generally, to determine the weight percentage of the albumin in the
nanoparticles, the amount of the albumin on the nanoparticles and
the total weight of the nanoparticles can be determined. The amount
of the albumin on the nanoparticles can be determined by, for
example, chromatography, such as reversed-phase chromatography,
size-exclusion chromatography and/or HPLC size-exclusion
chromatography methods, spectrophotometric measurements, or mass
spectrometric measurements. In some embodiments, the method
comprises separating the nanoparticles from the non-nanoparticle
portion by ultracentrifugation or gel filtration chromatography,
followed by analyzing the amount of the albumin on the
nanoparticles by, for example, size-exclusion chromatography.
Spectrophotometric measurements can be used to determine the amount
of the albumin on the nanoparticles. In some embodiments, the
weight percentage of the albumin in the nanoparticles is determined
upon reconstitution of the pharmaceutical composition. In some
embodiments, the weight percentage of the albumin in the
nanoparticles is determined upon storage of the pharmaceutical
composition.
In some embodiments, the total weight of the nanoparticles is
determined by addition of the amount of the albumin on the
nanoparticles and the amount of poorly water soluble drug (such as
rapamycin) in the nanoparticle. Amount of the poorly water soluble
drug (such as rapamycin) in the nanoparticles can be determined by,
for example, chromatography, such as reversed-phase high
performance liquid chromatography (RP-HPLC), spectrophotometric
measurements, or mass spectrometric measurements. In some
embodiments, the method comprises determining the amount of the
rapamycin in the nanoparticles, for example, by reversed-phase
HPLC.
In some embodiments, the amount of the albumin on the nanoparticles
and the amount of the rapamycin in the nanoparticles are used to
determine the total weight of the nanoparticles. The weight
percentage of the albumin in the nanoparticles can be calculated
from the amount of the albumin on the nanoparticles and the total
weight of the nanoparticles.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the weight percentage
of the albumin in the nanoparticles is about 15% to about 30% (such
as about 20% to about 25%, about 15% to about 24%, or about 15% to
about 20%). In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
weight percentage of the albumin in the nanoparticles is about any
one of 10%, 12%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%,
19%, 19.5%, 20%, 22%, 25%, 30%, 35%, or 40%. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if the weight percentage of the albumin in the
nanoparticles is about any one of 10-12%, 12-14%, 14-15%, 15-16%,
16-17%, 17-18%, 18-19%, 19-20%, 20-22%, 22-25%, 10-15%, 15-20%,
20-25%, 25-30%, 15-17%, 17-19%, 15-15.5%, 15.5-16%, 16-16.5%,
16.5-17%, 17-17.5%, 17.5-18%, 18-18.5%, 18.5-19%, 19-19.5%,
19.5-20%, 15.5-16.5%, 16.5-17.5%, 17.5-18.5%, 18.5-19.5%, 15-16.5%,
16-17.5%, 17-18.5%, 18-19.5%, 16.5-19%, 17.5-20%, 15-17.5%,
17.5-20%, 20-22.5%, or 22.5-24%.
Determination of Weight Ratio of Albumin to Poorly Water Soluble
Drug on the Nanoparticles
The methods of the present application in some embodiments require
determination of the weight ratio of the albumin on the
nanoparticles to the poorly water soluble drug (such as rapamycin)
in the nanoparticles.
Exemplary means for determining the amount of the albumin on the
nanoparticles and the amount of the poorly water soluble drug (such
as rapamycin) in the nanoparticles are discussed above. In some
embodiments, the weight ratio of albumin to poorly water soluble
drug (such as rapamycin) in the nanoparticles is determined by the
amount of the albumin on the nanoparticles over the amount of the
poorly water soluble drug (such as rapamycin) in the nanoparticles.
In some embodiment, the weight ratio of albumin to poorly water
soluble drug (such as rapamycin) in the nanoparticles is determined
upon storage of the pharmaceutical composition.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the weight ratio of
albumin to poorly water soluble drug (such as rapamycin) in the
nanoparticles is about 1:2 to about 1:6. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the weight ratio of albumin to poorly water soluble
drug (such as rapamycin) in the nanoparticles is about any of 1:1,
1:2, 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, 1:8, 1:9,
or 1:10. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
weight ratio of albumin to poorly water soluble drug (such as
rapamycin) in the nanoparticles is any of about 1:1 to about 1:2,
about 1:2 to about 1:3, about 1:3 to about 1:3.5, about 1:3.5 to
about 1:4, about 1:4 to about 1:4.5, about 1:4.5 to about 1:5,
about 1:5 to about 1:5.5, about 1:5.5 to about 1:6, about 1:6 to
about 1:6.5, about 1:6.5 to about 1:7, about 1:7 to about 1:8,
about 1:8 to about 1:9, about 1:9 to about 1:10, about 1:1 to about
1:4, about 1:4 to about 1:6, about 1:6 to about 1:10, about 1:3 to
about 1:4, about 1:4 to about 1:5, about 1:5 to about 1:6, about
1:6 to about 1:7, about 1:3.5 to about 1:4.5, about 1:4.5 to about
1:5.5, about 1:5.5 to about 1:6.5, or about 1:2 to about 1:6.
Determination of Rapamycin Concentration
The methods of present application in some embodiments require
determination of the concentration of rapamycin in the nanoparticle
portion of the composition.
The concentration of rapamycin in the nanoparticle portion of the
composition (such as a pharmaceutical composition) can be
determined by a variety of techniques including an HPLC assay using
UV absorbance. Briefly, for example, the nanoparticle portion of
composition (such as a pharmaceutical composition) is separated
from the non-nanoparticle portion of the composition by
ultracentrifugation, for example, at 50,000 rpm for 41 minutes at
25.degree. C. The supernatant is removed and the pellet is gently
washed with water twice. The pellet is then dispersed in a volume
of 50:50 acetonitrile:water solution, for example 3.0 ml, by
sonication. The sample is further diluted to ensure a homogenous
solution is formed. The sample is analyzed on an HPLC system
equipped with, for example, a Phenomenex, Curosil PFP guard column
(4.6 mm.times.30 mm, 5 .mu.m particle size) and a Phenomenex,
Curosil PFP analytical column (4.6 mm.times.250 mm, 5 .mu.m
particle size), a UV absorbance detector, and data acquisition.
Chromatograms are generated with the UV absorbance detector set at
228 nm. Comparison to analysis of rapamycin standards is used to
determine the concentration of rapamycin in the nanoparticle
portion of the composition. In some embodiments, the rapamycin
concentration in the nanoparticles is determined by the amount of
rapamycin in the nanoparticle portion of the composition in the
same volume of the original sample.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the rapamycin
concentration in the composition (such as a pharmaceutical
composition) is about any of about 2 mg/ml, 3 mg/ml, 4 mg/ml, 5
mg/ml, 6 mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, or 10 mg/ml.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the rapamycin
concentration in the composition (such as a pharmaceutical
composition) is any of about 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6
mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, or 10 mg/ml. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if the rapamycin concentration in the composition
(such as a pharmaceutical composition) is about 4.4-4.5 mg/ml,
4.5-4.6 mg/ml, 4.6-4.7 mg/ml, 4.7-4.8 mg/ml, 4.8-4.9 mg/ml, or
4.9-5.0 mg/ml. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
rapamycin in the suspension is about 5 mg/ml. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if the rapamycin in the composition (such as a
pharmaceutical composition) is about 5 mg/ml.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the rapamycin in the
nanoparticle portion of the composition (such as a pharmaceutical
composition) is any of about 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6
mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, or 10 mg/ml. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if the rapamycin in the nanoparticle portion of the
composition (such as a pharmaceutical composition) is about 4.2-4.3
mg/ml, 4.3-4.4 mg/ml, 4.4-4.5 mg/ml, 4.5-4.6 mg/ml, 4.6-4.7 mg/ml,
4.7-4.8 mg/ml, 4.9-5.0 mg/ml.
Determination of Nanoparticle Morphology
The methods of the present application in some embodiments require
determination of the nanoparticle morphology. The morphology of the
nanoparticles in an albumin-based nanoparticle composition can
affect particle solubility, dissolution rate, and disintegration
kinetics.
In some embodiments, the methods comprise determining the shape of
the nanoparticles, for example by microscopic methods such as
cryo-TEM. In some embodiments, the methods comprise determining the
thickness of the albumin coating on the nanoparticles, for example
by microscopic methods such as cryo-TEM. In some embodiments, the
methods comprise determining both the shape of the nanoparticles
and the thickness of the albumin coating on the nanoparticles by
microscopic methods, such as cryo-TEM. In some embodiments, the
shape of the nanoparticles is determined upon reconstitution of the
pharmaceutical composition. In some embodiments, the shape of the
nanoparticles is determined upon storage of the pharmaceutical
composition.
For example, the composition (such as a pharmaceutical composition)
can be rapidly cooled to cryogenic temperatures following
reconstitution of the composition (such as a pharmaceutical
composition) to form a vitreous form of the reconstituted
composition which can then be analyzed. The nanoparticles of the
composition (such as a pharmaceutical composition) remain in their
native structure during cryo-TEM sample preparation and image
recording. In some embodiments, cryo-TEM records the native
structure of the nanoparticles of the composition (such as a
pharmaceutical composition).
In some embodiments, the thickness of the albumin coating on the
nanoparticles is calculated based on measured parameters of the
nanoparticles, including for example the albumin-to-rapamycin ratio
of the nanoparticles. In some embodiments, the thickness of the
albumin coating on the nanoparticles is determined upon
reconstitution of the pharmaceutical composition. In some
embodiments, the thickness of the albumin coating on the
nanoparticles is determined upon storage of the pharmaceutical
composition.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the nanoparticles are
of irregular shape. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
nanoparticles have a non-smooth surface. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the nanoparticles are of irregular shape and have a
non-smooth surface. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
nanoparticles have a high degree of rugosity. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use of the nanoparticles are of irregular shape and
have a high degree of rugosity.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the thickness of the
albumin coating on the nanoparticles is about 5 nanometers to about
7 nanometers as measured by cryo-TEM. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the nanoparticles are of irregular shape and have an
albumin coating with a thickness of about 5 nanometers to about 7
nanometers as measured by cryo-TEM. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the albumin coating has a thickness of about any of
3 nanometers, 4 nanometers, 5 nanometers, 5.5 nanometers, 6
nanometers, 6.5 nanometers, 7 nanometers, 8 nanometers, or 9
nanometers as measured by cryo-TEM. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the albumin coating has a thickness of about any of
3-4 nanometers, 4-5 nanometers, 5-6 nanometers, 6-7 nanometers, 7-8
nanometers, 8-9 nanometers, 3-5 nanometers, 5-7 nanometers, 7-9
nanometers, 5-5.5 nanometers, 5.5-6 nanometers, 6-6.5 nanometers,
6.5-7 nanometers, 4.5-5.5 nanometers, 5.5-6.5 nanometers, 6.5-7.5
nanometers, 5-6.5 nanometers, or 5.5-7 nanometers as measured by
cryo-TEM.
In some embodiments, the methods described herein further comprise
determining the surface-to-volume ratio of the nanoparticles.
Surface-to-volume ratios of the nanoparticles can be determined,
for example, by microscopy methods, such as, cryo-TEM, atomic force
microscopy, or Fourier transform infrared spectroscopy. In some
embodiments, the surface-to-volume ratio of the nanoparticles is
determined upon reconstitution of the pharmaceutical composition.
In some embodiments, the surface-to-volume ratio of the
nanoparticles is determined upon storage of the pharmaceutical
composition.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the surface-to-volume
ratio of the nanoparticles is more than about 46.2:1 .mu.m.sup.-1,
or the surface-to-volume ratio of a perfect sphere having the same
particle size as the nanoparticles. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the surface-to-volume ratio of the nanoparticles is
more than about any of 30:1 .mu.m.sup.-1, 35:1 .mu.m.sup.-1, 40:1
.mu.m.sup.-1, 45:1 .mu.m.sup.-1, 46.2:1 .mu.m.sup.-1, 47:1
.mu.m.sup.-1, 48:1 .mu.m.sup.-1, 49:1 .mu.m.sup.-1, 50:1
.mu.m.sup.-1, 52:1 .mu.m.sup.-1, 55:1 .mu.m.sup.-1, 58:1
.mu.m.sup.-1, 60:1 .mu.m.sup.-1, 65:1 .mu.m.sup.-1, 70:1
.mu.m.sup.-1, 75:1 .mu.m.sup.-1, 80:1 .mu.m.sup.-1, 90:1
.mu.m.sup.-1, 100:1 .mu.m.sup.-1, 110:1 .mu.m.sup.-1, 120:1
.mu.m.sup.-1, 130:1 .mu.m.sup.-1, 140:1 .mu.m.sup.-1, 150:1
.mu.m.sup.-1, 160:1 .mu.m.sup.-1, 170:1 .mu.m.sup.-1, 180:1
.mu.m.sup.-1, 190:1 .mu.m.sup.-1, 200:1 .mu.m.sup.-1, 210:1
.mu.m.sup.-1, 220:1 .mu.m.sup.-1, 250:1 .mu.m.sup.-1, 300:1
.mu.m.sup.-1, or 400:1 .mu.m.sup.-1. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the surface-to-volume ratio of the nanoparticles is
more than any of about 30:1 .mu.m.sup.-1 to about 35:1
.mu.m.sup.-1, about 35:1 .mu.m.sup.-1 to about 40:1 .mu.m.sup.-1,
about 40:1 .mu.m.sup.-1 to about 45:1 .mu.m.sup.-1, about 46.2:1
.mu.m.sup.-1 to about 47:1 .mu.m.sup.-1, about 47:1 .mu.m.sup.-1 to
about 48:1 .mu.m.sup.-1, about 48:1 .mu.m.sup.-1 to about 49:1
.mu.m.sup.-1, about 49:1 .mu.m.sup.-1 to about 50:1 .mu.m.sup.-1,
about 50:1 .mu.m.sup.-1 to about 52:1 .mu.m.sup.-1, about 52:1
.mu.m.sup.-1 to about 55:1 .mu.m.sup.-1, about 55:1 .mu.m.sup.-1 to
about 58:1 .mu.m.sup.-1, about 58:1 .mu.m.sup.-1 to about 60:1
.mu.m.sup.-1, about 60:1 .mu.m.sup.-1 to about 65:1 .mu.m.sup.-1,
about 65:1 .mu.m.sup.-1 to about 70:1 .mu.m.sup.-1, about 70:1
.mu.m.sup.-1 to about 75:1 .mu.m.sup.-1, about 75:1 .mu.m.sup.-1 to
about 80:1 .mu.m.sup.-1, about 46.2:1 .mu.m.sup.-1 to about 50:1
.mu.m.sup.-1, about 50:1 .mu.m.sup.-1 to about 60:1 .mu.m.sup.-1,
about 60:1 .mu.m.sup.-1 to about 70:1 .mu.m.sup.-1, about 70:1
.mu.m.sup.-1 to about 80:1 .mu.m.sup.-1, about 46.2:1 .mu.m.sup.-1
to about 60:1 .mu.m.sup.-1, about 60:1 .mu.m.sup.-1 to about 80:1
.mu.m.sup.-1, about 50:1 .mu.m.sup.-1 to about 70:1 .mu.m.sup.-1,
about 48:1 .mu.m.sup.-1 to about 52:1 .mu.m.sup.-1, about 52:1
.mu.m.sup.-1 to about 65:1 .mu.m.sup.-1, about 65:1 .mu.m.sup.-1 to
about 80:1 .mu.m.sup.-1, about 80:1 .mu.m.sup.-1 to about 90:1
.mu.m.sup.-1, about 100:1 .mu.m.sup.-1 to about 120:1 .mu.m.sup.-1,
about 120:1 .mu.m.sup.-1 to about 140:1 .mu.m.sup.-1, about 140:1
.mu.m.sup.-1 to about 160:1 .mu.m.sup.-1, about 160:1 .mu.m.sup.-1
to about 180:1 .mu.m.sup.-1, about 180:1 .mu.m.sup.-1 to about
200:1 .mu.m.sup.-1, about 200:1 .mu.m.sup.-1 to about 220:1
.mu.m.sup.-1, about 220:1 .mu.m.sup.-1 to about 250:1 .mu.m.sup.-1,
about 250:1 .mu.m.sup.-1 to about 300:1 .mu.m.sup.-1, about 300:1
.mu.m.sup.-1 to about 400:1 .mu.m.sup.-1, about 30:1 .mu.m.sup.-1
to about 40:1 .mu.m.sup.-1, about 30:1 .mu.m.sup.-1 to about 45:1
.mu.m.sup.-1, about 80:1 .mu.m.sup.-1 to about 120:1 .mu.m.sup.-1,
about 120:1 .mu.m.sup.-1 to about 150:1 .mu.m.sup.-1, or about
150:1 .mu.m.sup.-1 to about 200:1 .mu.m.sup.-1.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the surface-to-volume
ratio of the nanoparticles is more than about 6/d, wherein d is the
average diameter of the nanoparticles (i.e. the surface-to-volume
ratio of a perfect sphere having the same particle size as the
nanoparticles). In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
surface-to-volume ratio of the nanoparticles is more than about any
of 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2,
2.3, 2.4, 2.5, 3, 4, 5, or more than 5 times of 6/d, wherein d is
the average diameter of the nanoparticles.
The surface-to-volume ratio of the nanoparticles is related to the
average diameter of the nanoparticles. As used herein, "diameter of
the nanoparticle" refers to the diameter of the sphere that has the
same volume or weight as the nanoparticle. "Average diameter of the
nanoparticles" is the average of the diameters of all nanoparticles
in the composition (such as a pharmaceutical composition). For
example, in some embodiments, when the average diameter of the
nanoparticles is no more than about 130 nm, the composition (such
as a pharmaceutical composition) is suitable for medical use if the
surface-to-volume ratio of the nanoparticles is more than about
46.2:1 .mu.m.sup.-1. In some embodiments, when the average diameter
of the nanoparticles is no more than about 130 nm, the composition
(such as a pharmaceutical composition) is suitable for medical use
if the surface-to-volume ratio of the nanoparticles is more than
about any of 46.2:1 .mu.m.sup.-1, 47:1 .mu.m.sup.-1, 48:1
.mu.m.sup.-1, 49:1 .mu.m.sup.-1, 50:1 .mu.m.sup.-1, 52:1
.mu.m.sup.-1, 55:1 .mu.m.sup.-1, 58:1 .mu.m.sup.-1, 60:1
.mu.m.sup.-1, 65:1 .mu.m.sup.-1, 70:1 .mu.m.sup.-1, 75:1
.mu.m.sup.-1, 80:1 m.sup.-1, 90:1 .mu.m.sup.-1, 100:1 .mu.m.sup.-1,
110:1 .mu.m.sup.-1, 120:1 .mu.m.sup.-1, or 140:1 .mu.m.sup.-1. In
some embodiments, when the average diameter of the nanoparticles is
no more than about 130 nm, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
surface-to-volume ratio of the nanoparticles is more than any of
about 46.2:1 .mu.m.sup.-1 to about 47:1 .mu.m.sup.-1, about 47:1
.mu.m.sup.-1 to about 48:1 .mu.m.sup.-1, about 48:1 .mu.m.sup.-1 to
about 49:1 .mu.m.sup.-1, about 49:1 .mu.m.sup.-1 to about 50:1
.mu.m.sup.-1, about 50:1 .mu.m.sup.-l to about 52:1 .mu.m.sup.-1,
about 52:1 .mu.m.sup.-1 to about 55:1 .mu.m.sup.-1, about 55:1
.mu.m.sup.-1 to about 58:1 .mu.m.sup.-1, about 58:1 .mu.m.sup.-1 to
about 60:1 .mu.m.sup.-1, about 60:1 .mu.m.sup.-1 to about 65:1
.mu.m.sup.-1, about 65:1 .mu.m.sup.-1 to about 70:1 .mu.m.sup.-1,
about 70:1 .mu.m.sup.-1 to about 75:1 .mu.m.sup.-1, about 75:1
.mu.m.sup.-1 to about 80:1 .mu.m.sup.-1, about 46.2:1 .mu.m.sup.-1
to about 50:1 .mu.m.sup.-1, about 50:1 .mu.m.sup.-1 to about 60:1
.mu.m.sup.-1, about 60:1 .mu.m.sup.-1 to about 70:1 .mu.m.sup.-1,
about 70:1 .mu.m.sup.-1 to about 80:1 .mu.m.sup.-1, about 80:1
.mu.m.sup.-1 to about 90:1 .mu.m.sup.-1, about 90:1 .mu.m.sup.-1 to
about 120:1 .mu.m.sup.-1, about 120:1 .mu.m.sup.-1 to about 140:1
.mu.m.sup.-1, about 46.1:1 .mu.m.sup.-1 to about 60:1 .mu.m.sup.-1,
about 60:1 .mu.m.sup.-1 to about 80:1 .mu.m.sup.-1, about 50:1
.mu.m.sup.-1 to about 70:1 .mu.m.sup.-1, about 48:1 .mu.m.sup.-1 to
about 52:1 .mu.m.sup.-1, about 52:1 .mu.m.sup.-1 to about 65:1
.mu.m.sup.-1, or about 65:1 .mu.m.sup.-1 to about 80:1
.mu.m.sup.-1. In some embodiments, when the average diameter of the
nanoparticles is about 60 nm to about 190 nm, the composition (such
as a pharmaceutical composition) is suitable for medical use if the
surface-to-volume ratio of the nanoparticles is more than about any
of 31.5:1 .mu.m.sup.-1, 35:1 .mu.m.sup.-1, 40:1 .mu.m.sup.-1, 45:1
.mu.m.sup.-1, 50:1 .mu.m.sup.-1, 55:1 .mu.m.sup.-1, 60:1
.mu.m.sup.-1, 65:1 .mu.m.sup.-1, 70:1 .mu.m.sup.-1, 80:1
.mu.m.sup.-1, 90:1 .mu.m.sup.-1, 100:1 .mu.m.sup.-1, 110:1
.mu.m.sup.-1, 120:1 .mu.m.sup.-1, 130:1 .mu.m.sup.-1, 140:1
.mu.m.sup.-1, 150:1 .mu.m.sup.-1, 160:1 .mu.m.sup.-1, 170:1
.mu.m.sup.-1, 180:1 .mu.m.sup.-1, 190:1 .mu.m.sup.-1, 200:1
.mu.m.sup.-1, or more than 200:1 .mu.m.sup.-1. In some embodiments,
when the average diameter of the nanoparticles is about 120 nm to
about 140 nm, the composition (such as a pharmaceutical
composition) is suitable for medical use if the surface-to-volume
ratio of the nanoparticles is more than about any of 42.9:1
.mu.m.sup.-1, 45:1 .mu.m.sup.-1, 50:1 .mu.m.sup.-1, 55:1
.mu.m.sup.-1, 60:1 .mu.m.sup.-1, 65:1 .mu.m.sup.-1, 70:1
.mu.m.sup.-1, 75:1 .mu.m.sup.-1, 80:1 .mu.m.sup.-1, 85:1
.mu.m.sup.-1, 90:1 .mu.m.sup.-1, 95:1 .mu.m.sup.-1, 100:1
.mu.m.sup.-1 or more than 100:1 .mu.m.sup.-1.
Determination of Particle Size and Polydispersity
The methods of the present application in some embodiments require
determination of the size of nanoparticles in the composition (such
as a pharmaceutical composition). Particle size impacts the
dissolution rate of nanoparticles, controls the solubility of
nanoparticles, and contributes to the functional behavior of the
nanoparticles.
In some embodiments, the methods comprise determining the size of
the nanoparticles in the pharmaceutical composition. In some
embodiments, the methods comprise determining the polydispersity
index of the nanoparticles in the pharmaceutical composition. In
some embodiments, the methods comprise determining the size
distribution of the nanoparticles in the pharmaceutical
composition. In some embodiments, the polydispersity index of the
nanoparticles is determined upon reconstitution of the
pharmaceutical composition. In some embodiments, the polydispersity
index of the nanoparticles is determined upon storage of the
pharmaceutical composition.
In some embodiments, the particle size is determined by laser
diffraction techniques, such as dynamic light scattering. In some
embodiments, the size is determined by volume weighted arithmetic
mean particle diameter (D4,3) using a laser diffraction technique.
In some embodiments, the particle size is determined by disc
centrifugation methods. In some embodiments, the particle size is
determined by tunable resistive pulse sensing (TRPS). In some
embodiments, the particle size is determined by laser diffraction
polarization intensity differential scattering (PIDS-LD). In some
embodiments, the particle size is determined by sucrose gradient
centrifugation. In some embodiments, the particle size is
determined by analytical centrifugation.
In some embodiments, the polydispersity index is determined by, for
example, dynamic light scattering. Dv.sub.50 is the volume-weighted
median particle diameter.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the average particle
size of the nanoparticles in the pharmaceutical composition is less
than about 200 nm. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
average particle size of the nanoparticles in the pharmaceutical
composition is less than about any of 260 nm, 240 nm, 220 nm, 200
nm, 180 nm, 160 nm, 140 nm, 120 nm, 100 nm, 80 nm, or 60 nm. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the average particle
size of the nanoparticles in the pharmaceutical composition is
about any of 60-80 nm, 80-100 nm, 100-120 nm, 120-140 nm, 140-160
nm, 160-180 nm, 180-200 nm, 200-220 nm, 220-240 nm, 240-260 nm,
80-120 nm, 120-160 nm, 160-200 nm, 200-240 nm, 50-100 nm, 100-150
nm, 150-200 nm, 200-250 nm, 100-105 nm, 105-115 nm, 115-125 nm,
125-135 nm, 135-145 nm, 145-155 nm, 155-160 nm, 100-110 nm, 110-120
nm, 120-130 nm, 130-140 nm, 140-150 nm, 150-160 nm, 105-125 nm,
125-145 nm, 145-160 nm, 100-130 nm, 130-160 nm, 105-135 nm, 135-160
nm, 100-140 nm, 120-160 nm, 110-150 nm, 100-150 nm, 105-155 nm, or
100-160 nm. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
average particle size of the nanoparticles in the pharmaceutical
composition is about 130 nm.
The parameter ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) describes the span
of distribution of the particle sizes of the nanoparticles.
Dv.sub.50 refers to the volume-weighted median particle diameter.
Dv.sub.90 refers to the particle diameter where 90% of the volume
of all nanoparticles is contained in nanoparticles with smaller
diameters. Dv.sub.10 refers to the particle diameter where 10% of
the volume of all nanoparticles is contained in nanoparticles with
smaller diameters. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
nanoparticles in the pharmaceutical composition have a span of size
distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of about 0.8 to
about 1.5. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
nanoparticles in the pharmaceutical composition have a span of size
distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of about any of 0.5,
0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, or 1.8.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the nanoparticles in
the pharmaceutical composition have a size distribution
((DV.sub.90-DV.sub.10/DV.sub.50)) of about any of 0.5-0.6, 0.6-0.7,
0.7-0.8. 0.8-0.9, 0.9-1, 1-1.1, 1.1-1.2, 1.2-1.3, 1.3-1.4, 1.4-1.5,
1.5-1.6, 1.6-1.7, 1.7-1.8, 0.9-1.1, 1.1-1.3, 1.3-1.5, 1.5-1.7,
0.6-0.8, 0.8-1, 1-1.2, 1.2-1.4, 1.4-1.6, 1.6-1.8, 0.5-0.8, 0.8-1.1,
1.1-1.4, 1.4-1.8, 0.8-1.1, 1.1-1.4, 0.9-1.2, 1.2-1.5, 0.8-1.2,
0.9-1.3, 1-1.4, 1.1-1.5, 0.8-1.3, 0.9-1.4, 1-1.5, 0.8-1.4, 0.9-1.5,
or 0.8-1.5.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the nanoparticles in
the pharmaceutical composition have a polydispersity index of less
than about 0.3. In some embodiments, the nanoparticles in the
pharmaceutical composition have a polydispersity index of less than
about any of 0.3, 0.25, 0.2, 0.15, 0.1, or 0.05. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the nanoparticles in the
pharmaceutical composition have a polydispersity index of about any
of 0.05-0.07, 0.07-0.09, 0.09-0.11, 0.11-0.13, 0.13-0.15,
0.15-0.17, 0.17-0.2, 0.2-0.25, 0.25-0.3, 0.05-0.09, 0.09-0.13,
0.13-0.17, 0.17-0.25, 0.06-0.08, 0.08-0.12, 0.12-0.16, 0.16-0.18,
0.18-0.22, 0.22-0.28, 0.28-0.3, 0.06-0.12, 0.12-0.18, 0.18-0.3,
0.05-0.1, 0.1-0.15, 0.15-0.2, or 0.2-0.3.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) has a size distribution curve
similar to that of ABI-009. For details regarding ABI-009, See
Gonzalez-Angulo, A. M. et al. Clin. Cancer Res. 2013.
Determination of Surface Potential
The methods of the present application in some embodiments comprise
determining the surface potential, such as zeta-potential, of the
nanoparticles. Particle surface potential, such as zeta-potential,
can play an important role in preventing the particles from
aggregating.
Zeta-potential of the nanoparticles can be determined by
techniques, such as, microelectrophoresis, electrophoretic light
scattering, or dynamic electrophoretic mobility. In some
embodiments, the zeta-potential of the nanoparticles can be
determined by tunable resistive pulse sensing (TRPS). In some
embodiments, the zeta-potential of the nanoparticles is determined
upon reconstitution of the pharmaceutical composition. In some
embodiments, the zeta-potential of the nanoparticles is determined
upon storage of the pharmaceutical composition.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the nanoparticles in
the pharmaceutical composition have a zeta-potential of about -20
mV to about -35 mV. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
nanoparticles in the pharmaceutical composition have a
zeta-potential of about -25 mV. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the nanoparticles in the pharmaceutical composition
have a zeta-potential of about any of -40 mV, -35 mV, -30 mV, -29
mV, -28 mV, -27 mV, -26 mV, -25 mV, -24 mV, -23 mV, -22 mV, -21 mV,
-20 mV, -15 mV, -10 mV. In some embodiments, the composition (such
as a pharmaceutical composition) is suitable for medical use if the
nanoparticles in the pharmaceutical composition have a
zeta-potential of any of about -40 mV to about -35 mV, about -35 mV
to about -30 mV, about -30 mV to about -25 mV, about -25 mV to
about -20 mV, about -20 mV to about -15 mV, about -15 mV to about
-10 mV, about -30 mV to about -28 mV, about -28 mV to about -26 mV,
about -26 mV to about -24 mV, about -24 mV to about -22 mV, about
-22 mV to about -20 mV, about -29 mV to about -27 mV, about -27 mV
to about -25 mV, about -25 mV to about -23 mV, about -23 mV to
about -21 mV, about -30 mV to about -26 mV, about -26 mV to about
-22 mV, about -28 mV to about -24 mV, about -24 mV to about -20 mV,
about -30 mV to about -25 mV, about -25 mV to about -20 mV, or
about 30 mV to about -20 mV.
Determination of Drug Crystallinity
The methods of the present application in some embodiments comprise
determining the crystalline state of the poorly water soluble drug
(such as rapamycin) in the composition (such as a pharmaceutical
composition). In some embodiments, the method comprises determining
the crystalline state of rapamycin by X-ray diffraction. In some
embodiments, the method comprises determining the crystallinity of
rapamycin by light microscopy, such as polarized light microscopy.
In some embodiments, the method comprises determining the
crystallinity of the poorly water soluble drug (such as rapamycin)
by both X-ray diffraction and a polarized light microscopy method.
In some embodiments, the method comprises determining the
crystallinity of the poorly water soluble drug (such as rapamycin)
by Raman spectroscopy. In some embodiments, the method comprises
determining the crystallinity of the poorly water soluble drug
(such as rapamycin) by second harmonic generation microscopy. In
some embodiments, the method comprises determining the
crystallinity of the poorly water soluble drug (such as rapamycin)
by X-ray powder diffraction. In some embodiments, the method
comprises determining the crystallinity of the poorly water soluble
drug (such as rapamycin) by differential scanning calorimetry. In
some embodiments, the method comprises determining the
crystallinity of the poorly water soluble drug (such as rapamycin)
by thermal gravimetric analysis. In some embodiments, the method
comprises determining the crystallinity of the poorly water soluble
drug (such as rapamycin) using one or more technique selected from
the group consisting of X-ray diffraction, X-ray powder
diffraction, light microscopy, polarized light microscopy, Raman
spectroscopy, second harmonic generation microscopy, differential
scanning calorimetry, and thermal gravimetric analysis.
In some embodiments, the method comprises determining the
crystallinity of the poorly water soluble drug (such as rapamycin)
by qualitatively determining one or more crystalline forms of the
poorly water soluble drug (such as rapamycin). In some embodiments,
the method comprises determining the crystallinity of the poorly
water soluble drug (such as rapamycin) by qualitatively determining
two crystalline forms of the poorly water soluble drug (such as
rapamycin). In some embodiments, the method comprises determining
the crystallinity of the poorly water soluble drug (such as
rapamycin) by quantitatively determining one or more crystalline
forms of the poorly water soluble drug (such as rapamycin). In some
embodiments, the method comprises determining the crystallinity of
the poorly water soluble drug (such as rapamycin) by quantitatively
determining two crystalline forms of the poorly water soluble drug
(such as rapamycin). In some embodiments, the method comprises
determining the crystallinity of the poorly water soluble drug
(such as rapamycin) by qualitatively and quantitatively determining
one or more crystalline forms of the poorly water soluble drug
(such as rapamycin). In some embodiments, the method comprises
determining the crystallinity of the poorly water soluble drug
(such as rapamycin) by qualitatively and quantitatively determining
two crystalline forms of the poorly water soluble drug (such as
rapamycin).
In some embodiments, the determination of rapamycin crystallinity
is determined immediately after reconstitution. In some
embodiments, the determination of rapamycin crystallinity is
determined after storage, for example after storage for at least
about any of 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 40
hours, 44 hours, 48 hours, 52 hours, 56 hours, 60 hours, 64 hours,
or 72 hours (for example at room temperature, under refrigerated
condition, or at about 40.degree. C.).
In some embodiments, nanoparticles are isolated by, for example,
ultracentrifugation or gel permeation chromatography. In some
embodiments, following ultracentrifugation, the supernatant is
decanted and the pellet is washed with water. The isolated
nanoparticles are then dried by, for example, lyophilization.
In some embodiments, subsequent analysis of the nanoparticles by
X-ray diffraction can determine the physical state of rapamycin in
the nanoparticles. Non-crystalline rapamycin in the nanoparticles
will exhibit broad scattering halos, indicative of an amorphous
material (e.g., non-crystalline). Crystalline rapamycin in the
nanoparticles will exhibit numerous well-defined scattering
peaks.
In some embodiments, X-ray powder diffraction of the dried sample
is used (alone or in addition to any other method described herein)
to determine the physical state of the rapamycin in the
nanoparticles. Non-crystalline rapamycin in the nanoparticles will
exhibit broad scattering halos, indicative of an amorphous material
(e.g., non-crystalline). Crystalline rapamycin in the nanoparticles
will exhibit numerous well-defined scattering peaks such as sharp
scattering peaks.
In some embodiments, polarized light microscopy of a reconstituted
suspension of nanoparticles is used (alone or in addition to any
other method described herein) to determine the physical state of
rapamycin in the nanoparticles. A birefringence test can be
performed with an optical microscope to determine if the rapamycin
in the nanoparticles is crystalline or non-crystalline. Absence of
birefringence indicates that the rapamycin remained amorphous.
In some embodiments, Raman spectroscopy can be used (alone or in
addition to any other method described herein) to determine the
physical state of the rapamycin in the nanoparticles. Crystalline
rapamycin, such as crystalline rapamycin hydrate, has peaks at
945-947 cm.sup.-1, 1320 cm.sup.-1, 1349 cm.sup.-1, 1360 cm.sup.-1,
1631-1647 cm.sup.-1 and a shoulder on the peak at 1715
cm.sup.-1.
In some embodiments, differential scanning calorimetry can be used
(alone or in addition to any other method described herein) to
determine the physical state of the rapamycin in the nanoparticles.
The temperature of thermal transition, e.g., glass transition, can
be used to determine the presence of crystalline rapamycin in the
nanoparticles. For example, amorphous rapamycin in the
nanoparticles has a thermal transition of 150.degree. C., the
crystalline melt of the anhydrous crystal is 213.degree. C., and
the solid-state transition in the crystal hydrate is 166.degree.
C.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the rapamycin in the
pharmaceutical composition is non-crystalline. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if the rapamycin in the reconstituted
pharmaceutical composition is non-crystalline for at least about
any of 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 40 hours,
44 hours, 48 hours, 52 hours, 56 hours, 60 hours, 64 hours, or 72
hours. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
paclitaxel in the lyophilized pharmaceutical composition is
non-crystalline upon storage at about 40.degree. C. for at least
about any of 1 month, 2 months, 3 months, 4 months, 5 months, 6
months, 12 months, 24 months, or more months. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if the rapamycin in the pharmaceutical composition
is non-crystalline upon storage of the composition (such as a
pharmaceutical composition) at about 4.degree. C. for at least
about 24 hours. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
rapamycin in the pharmaceutical composition is non-crystalline upon
storage of the composition (such as a pharmaceutical composition)
for at least about any of 6 hours, 12 hours, 18 hours, 24 hours, 36
hours, 40 hours, 44 hours, 48 hours, 52 hours, 56 hours, 60 hours,
64 hours, or 72 hours at, for example at room temperature, under
refrigerated condition, or at about 40.degree. C. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if no crystalline form of rapamycin can
be detected in the pharmaceutical composition. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if no crystalline form of rapamycin can be detected
at a limit of 2% of the total rapamycin mass in the pharmaceutical
composition. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if less
than about 0.01%, 0.008%, 0.005%, or 0.003% of the rapamycin in the
pharmaceutical composition is in a crystalline form. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if no crystalline form of rapamycin can
be detected in the pharmaceutical composition in solution. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if no crystalline form of rapamycin can
be detected at a limit of 2% of the total rapamycin mass in the
pharmaceutical composition in solution. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if less than 0.01%, 0.008%, 0.005%, or 0.003% of the
rapamycin in the pharmaceutical composition in solution is in a
crystalline form.
Determination of Distribution of the Total Albumin and the Total
Poorly Water Soluble Drug Between Nanoparticles and the
Non-Nanoparticle Portion
The methods of the present application in some embodiments comprise
determining the distribution of the total albumin and the total
poorly water soluble drug (such as rapamycin) between nanoparticles
and the non-nanoparticle portion of the composition (such as a
pharmaceutical composition). In some embodiments, the method
comprises determining the distribution of the total albumin between
the nanoparticles and the non-nanoparticle portion of the
composition (such as a pharmaceutical composition). In some
embodiments, the method comprises determining the distribution of
the total rapamycin between the nanoparticles and the
non-nanoparticle portion of the composition (such as a
pharmaceutical composition). In some embodiments, the distribution
of the total rapamycin between the nanoparticles and the
non-nanoparticle portion is determined upon reconstitution of the
pharmaceutical composition. In some embodiments, the distribution
of the total rapamycin between the nanoparticles and the
non-nanoparticle portion is determined upon storage of the
pharmaceutical composition.
The distribution of the total albumin between the nanoparticles and
the non-nanoparticle portion of the composition (such as a
pharmaceutical composition) can be determined by measuring the
amount of the albumin on the nanoparticles of the composition (such
as a pharmaceutical composition) and/or the amount of the albumin
in the non-nanoparticle portion of the composition (such as a
pharmaceutical composition). Exemplary methods for measuring the
amount albumin in the nanoparticles and the non-nanoparticle
portion are discussed above. In some embodiments, the distribution
of the total albumin is determined as a percentage of the total
albumin in the composition (such as a pharmaceutical composition)
in the non-nanoparticle portion of the composition (such as a
pharmaceutical composition).
In some embodiments, the distribution of the total rapamycin in the
nanoparticles and/or the non-nanoparticle portion of the
composition (such as a pharmaceutical composition) can be
determined by measuring the amount of the rapamycin in the
nanoparticles of the composition (such as a pharmaceutical
composition) and the amount of the rapamycin in the
non-nanoparticle portion of the composition (such as a
pharmaceutical composition). Exemplary methods for measuring the
amount of the rapamycin in the nanoparticles and the
non-nanoparticle portion are discussed above. In some embodiments,
the distribution of the total rapamycin is determined as a
percentage of the total rapamycin in the composition (such as a
pharmaceutical composition) in the nanoparticle portion of the
composition (such as a pharmaceutical composition).
The amount of the total rapamycin in the composition (such as a
pharmaceutical composition) associated with nanoparticles can be
determined by reversed-phase high performance liquid chromatography
(RP-HPLC). For example, the nanoparticles can first be isolated by
ultracentrifugation or gel filtration chromatography. Subsequently,
the amount of the rapamycin in the nanoparticles can then be
determined by assaying with quantitative RP-HPLC methods or mass
spectrometric methods. The amount of the rapamycin measured from
the isolated nanoparticles can then be compared with the amount of
the total rapamycin in the composition (such as a pharmaceutical
composition) to calculate the percentage of the total rapamycin in
the composition (such as a pharmaceutical composition) that is
associated with the nanoparticles. In some embodiments, the amount
of the total rapamycin in the composition (such as a pharmaceutical
composition) associated with the nanoparticles can be determined by
measuring the amount of the rapamycin not associated with the
nanoparticles. For example, following ultracentrifugation to pellet
the nanoparticles, the amount of rapamycin in the resulting
supernatant can be assayed by RP-HPLC methods to determine the
amount of rapamycin in solution (i.e., not associated with
nanoparticles). The amount of the rapamycin measured from the
supernatant and the amount of the total rapamycin in the
composition (such as a pharmaceutical composition) can be used to
calculate the percentage of the total rapamycin in the composition
(such as a pharmaceutical composition) that is associated with the
nanoparticles.
The quantity of the albumin in the nanoparticles can be determined
from isolated nanoparticles assayed for albumin content by
size-exclusion chromatography or HPLC size-exclusion chromatography
methods. The albumin in the non-nanoparticle portion can be
determined by assaying the supernatant using a similar
size-exclusion chromatography method.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if at least about 95% of
the total rapamycin in the composition (such as a pharmaceutical
composition) are associated with the nanoparticles. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if at least about any of 99.5%, 99%,
98.5%, 98%, 97.5%. 97%, 96.5%, 96%, 95%, 94%, 93%, 92%, 91%, 90%,
85%, or 80% of the total rapamycin in the composition (such as a
pharmaceutical composition) are associated with the nanoparticles.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if about any of 80-85%,
85-90%, 90-95%, 95-97%, 97-97.5%, 97.5-98%, 98-98.5%, 98.5-99%,
99-99.5%, 97-98%, 98-99%, 97.5-98.5%, 98.5-99.5%, 97-98.5%, 97-99%,
97-99.5% of the total rapamycin in the composition (such as a
pharmaceutical composition) are associated with the
nanoparticles.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if at least about 95% of
the total albumins in the composition (such as a pharmaceutical
composition) are in the non-nanoparticle portion of the composition
(such as a pharmaceutical composition). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if at least about any of 99.5%, 99%, 98.5%, 98%, 97.5%,
97%, 96.5%, 96%, 95.5%, 95%, 94%, 93%, 92%, 91%, 90%, 85%, or 80%
of the total albumin in the composition (such as a pharmaceutical
composition) are in the non-nanoparticle portion of the composition
(such as a pharmaceutical composition). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about any of 80-85%, 85-90%, 90-95%, 95-96%,
96-96.5%, 96.5-97%, 97-97.5%, 97.5-98%, 98-98.5%, 98.5-99%,
99-99.5%, 96-97%, 97-98%, 98-99%, 96.5-97.5%, 97.5-98.5%,
98.5-99.5%, 96-98%, 98-99.5%, 96-98.5%, 97-99.5%, or 96-99.5% of
the total albumin in the composition (such as a pharmaceutical
composition) are in the non-nanoparticle portion of the composition
(such as a pharmaceutical composition).
Determination of Albumin Oligomeric Status in the Composition
The methods of the present application in some embodiments comprise
determining the oligomeric status (e.g., polymers, monomers,
dimers, and/or oligomers) of the total albumin in the composition
(such as a pharmaceutical composition).
The oligomeric forms of the total albumin in the composition (such
as a pharmaceutical composition) (e.g., monomeric, dimeric,
oligomeric, polymeric) can be determined by, for example,
size-exclusion chromatography, such as gel permeation
chromatography or HPLC size-exclusion chromatography methods, or
polyacrylamide gel electrophoresis. For example, the oligomeric
status can be determined by analysis of the total albumin in the
composition (such as a pharmaceutical composition) by
size-exclusion chromatography. The different classes of albumins
can be determined based on differing retention time of albumin when
subject to a chromatography (such as size-exclusion
chromatography). The distribution of the components can be
confirmed by permeation chromatography. In some embodiments, the
oligomeric status of the total albumin in the pharmaceutical
composition is determined upon reconstitution of the pharmaceutical
composition. In some embodiments, the oligomeric status of the
total albumin in the pharmaceutical composition is determined upon
storage of the pharmaceutical composition.
The amount of monomeric albumin in the composition (such as a
pharmaceutical composition) can be compared with the amount of the
total albumin in the composition (such as a pharmaceutical
composition) to calculate the percentage of the total albumin in
the composition (such as a pharmaceutical composition) in the form
of monomers. The amount of dimeric albumin in the composition (such
as a pharmaceutical composition) can be compared with the amount of
the total albumin in the composition (such as a pharmaceutical
composition) to calculate the percentage of the total albumin in
the composition (such as a pharmaceutical composition) in the form
of dimers. The amount of oligomeric albumin in the composition
(such as a pharmaceutical composition) can be compared with the
amount of the total albumin in the composition (such as a
pharmaceutical composition) to calculate the percentage of the
total albumin in the composition (such as a pharmaceutical
composition) in the form of oligomers. The amount of polymeric
albumin in the composition (such as a pharmaceutical composition)
can be compared with the amount of the total albumin in the
composition (such as a pharmaceutical composition) to calculate the
percentage of the total albumin in the composition (such as a
pharmaceutical composition) in the form of polymers.
In some embodiments, the separation range for the size-exclusion
chromatography is about 10,000 daltons to about 500,000 daltons. In
some embodiments, the size-exclusion chromatography is run with a
TSKgel G3000 SWXL column. In some embodiments, the size-exclusion
chromatography is run with a column of TOSOH TSKgel G3000 SWXL,
7.8.times.300 mm, 5 .mu.m or equivalent. In some embodiments, the
size-exclusion chromatography is run with a flow rate of about 1
mL/min. In some embodiments, the size-exclusion chromatography is
run at ambient temperature. In some embodiments, the size-exclusion
chromatography is run with a column of TOSOH TSKgel G3000 SWXL,
7.8.times.300 mm, 5 .mu.m or equivalent, at a flow rate of about 1
mL/min at room temperature.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if at least about 2% of
the total albumin in the composition (such as a pharmaceutical
composition) is in the form of polymers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about 2% to about 5% of the total albumin in the
composition (such as a pharmaceutical composition) is in the form
of polymers. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if at least
about any of 1%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, or 8% of the
total albumin in the composition (such as a pharmaceutical
composition) is in the form of polymers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the percentage of polymeric albumin among the total
albumin in the pharmaceutical composition is about any of 1-2%,
2-2.5%, 2.5-3%, 3-3.5%, 3.5-4%, 4-4.5%, 4.5-5%, 5-6%, 6-8%, 2-3%,
3-4%, 4-5%, 5-6%, 2.5-3.5%, 3.5-4.5%, 4.5-5.5%, 2-4%, 4-6%, 3-5%,
2.5-4.5%, 4.5-6%, 2-3.5%, or 2-5%.
The present application in some embodiments provides albumin-based
rapamycin compositions (such as pharmaceutical compositions)
suitable for medical use in which about 75% to about 87% of the
total albumin in the composition (such as a pharmaceutical
composition) is in the form of monomers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about any of 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%,
82%, 83%, 84%, 85%, 86%, 87%, or 90% of the total albumin in the
composition (such as a pharmaceutical composition) is in the form
of monomers. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if about
any of 70-75%, 75-76%, 76-77%, 77-78%, 78-79%, 79-80%, 80-81%,
81-82%, 82-83%, 83-84%, 84-85%, 85-86%, 86-87%, 87-90%, 75-77%,
77-79%, 79-81%, 81-83%, 83-85%, 85-87%, 76-78%, 78-80%, 80-82%,
82-84%, 84-86%, 75-78%, 78-81%, 81-84%, 84-87%, 75-80%, 80-87%,
78-84%, 84-87%, 75-85%, 77-87%, or 75-87% of the total albumin in
the composition (such as a pharmaceutical composition) is in the
form of monomers.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if about 1% to about 4% of
the total albumin in the composition (such as a pharmaceutical
composition) is in the form of oligomers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about any of 0.5%, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%,
2.2%, 2.4%, 2.6%, 2.8%, 3%, 3.2%, 3.4%, 3.7%, 4%, or 4.5% of the
total albumin in the composition (such as a pharmaceutical
composition) is in the form of oligomers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about any of 0.5-1%, 1-1.2%, 1.2-1.4%, 1.4-1.6%,
1.6-1.8%, 1.8-2%, 2-2.2%, 2.2-2.4%, 2.4-2.6%, 2.6-2.8%, 2.8-3%,
3-3.2%, 3.2-3.4%, 3.4-3.7%, 3.7%-4%, 4-4.5%, 1-1.4%, 1.4-1.8%,
1.8-2.2%, 2.2-2.6%, 2.6-3%, 3-3.4%, 3.4-4%, 1.2-1.6%, 1.6-2%,
2-2.4%, 2.4-2.8%, 2.8-3.2%, 3.2-3.7%, 11.8%, 1.8%-2.4%, 2.4-3%,
3-3.7% 1.4-2%, 2-2.6%, 2.6-3.2%, 3.2-4%, 1-2%, 2-3%, 3-4%,
1.5-2.5%, 2.5-3.5%, 3.5-4%, 1-2.5%, 2.5-4%, or 1-4% of the total
albumin in the composition (such as a pharmaceutical composition)
is in the form of oligomers.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if about 6% to about 13%
of the total albumin in the composition (such as a pharmaceutical
composition) is in the form of dimers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about any of 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%,
12%, 13%, 14%, or 15% of the total albumin in the composition (such
as a pharmaceutical composition) is in the form of dimers. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if about any of 3-4%, 4-5%, 5-6%, 6-7%,
7-8%, 8-9%, 9-10%, 10-11%, 11-12%, 12-13%, 13-14%, 14-15%,
5.5-6.5%, 6.5-7.5%, 7.5-8.5%, 8.5-9.5%, 9.5-10.5%, 10.5-11.5%,
6-8%, 8-10%, 10-12%, 7-9%, 9-11%, 6.5-8.5%, 8.5-10.5%, 10.5-13%,
6-9%, 9-13%, 6-10%, 8-13%, or 6-13% of the total albumin in the
composition (such as a pharmaceutical composition) is in the form
of dimers.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if about 2% to about 5% of
the total albumin in the composition (such as a pharmaceutical
composition) is in the form of polymers, and about 75% to about 85%
of the total albumin in the composition (such as a pharmaceutical
composition) in the form of monomers. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if about any of 1%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%,
6%, or 8% of the total albumin in the composition (such as a
pharmaceutical composition) is in the form of polymers, and about
any of 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%,
86%, 87%, or 90% of the total albumin in the composition (such as a
pharmaceutical composition) is in the form of monomers. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if about any of 1-2%, 2-2.5%, 2.5-3%,
3-3.5%, 3.5-4%, 4-4.5%, 4.5-5%, 5-6%, 6-8%, 2-3%, 3-4%, 4-5%, 5-6%,
2.5-3.5%, 3.5-4.5%, 4.5-5.5% 2-4%, 4-6%, 3-5%, 2.5-4.5%, 4.5-6%,
2-3.5%, or 2-5% of the total albumin in the composition (such as a
pharmaceutical composition) is in the form of polymers, and about
any of 70-75%, 75-76%, 76-77%, 77-78%, 78-79%, 79-80%, 80-81%,
81-82%, 82-83%, 83-84%, 84-85%, 85-86%, 86-87%, 87-90%, 75-77%,
77-79%, 79-81%, 81-83%, 83-85%, 85-87%, 76-78%, 78-80%, 80-82%,
82-84%, 84-86%, 75-78%, 78-81%, 81-84%, 84-87%, 75-80%, 80-87%,
78-84%, 84-87%, 75-85%, 77-87%, or 75-87% of the total albumin in
the composition (such as a pharmaceutical composition) is in the
form of monomers.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the oligomeric status
of the total albumin in the composition (such as a pharmaceutical
composition) do not change significantly upon storage (such as at
about 25.degree. C. for about any of 3 months, 6 months, 9 months,
12 months, 18 months, 24 months, or 36 months). In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the percentage of albumin polymers
in the total albumin in the composition (such as a pharmaceutical
composition) does not increase by any of 5%, 10%, 20%, 30%, 40%,
50%, or more than 50% upon storage (such as at about 25.degree. C.
for 18 months). In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
percentage of albumin polymers in the total albumin in the
composition (such as a pharmaceutical composition) does not
decrease by any of 5%, 10%, 20%, 30%, 40%, 50%, or more than 50%
upon storage (such as at about 25.degree. C. for 18 months). In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the percentage of
albumin monomers in the total albumin in the composition (such as a
pharmaceutical composition) does not increase by any of 5%, 10%,
15%, 20%, 30%, 40%, 50%, or more than 50% upon storage (such as at
about 25.degree. C. for 18 months). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the percentage of albumin monomers in the total
albumin in the composition (such as a pharmaceutical composition)
does not decrease by any of 5%, 10%, 15%, 20%, 30%, 40%, 50%, or
more than 50% upon storage (such as at about 25.degree. C. for 18
months).
Determination of Recovery of Poorly Water Soluble Drug Following
Filtration
The methods of the present application in some embodiments comprise
determining the recovery of poorly water soluble drug (such as
rapamycin) following filtration. Loss of poorly water soluble drug
(such as rapamycin) following 0.2 micron filtration is a measure of
the faction of poorly water soluble drug (such as rapamycin) mass
associated with particles large than 200 nm. This measure can be
more sensitive to the large particle fraction than particle sizing
techniques.
In some embodiments, the methods comprise determining the recovery
of rapamycin following filtration with a 0.2 micron filter
immediately after reconstitution. In some embodiments, the method
comprises determining the recovery of rapamycin following
filtration with a 0.2 micron filter upon storage (after storage for
at least about any of 6 hours, 12 hours, 24 hours, 36 hours, 48
hours, 72 hours or more hours (for example under storage at room
temperature, under refrigerated conditions, or under accelerated
storage condition (for example at 40.degree. C.)).
The recovery of the rapamycin in the composition (such as a
pharmaceutical composition) following 0.2 micron filtration can be
determined by measuring the amount, such as weight, of rapamycin in
the composition (such as a pharmaceutical composition) that passes
through the 0.2 micron filter. As discussed above, the amount of
rapamycin can be measured by RP-HPLC techniques. In some
embodiments, to determine the recovery, the amount of rapamycin
that remains in the composition (such as a pharmaceutical
composition) following 0.2 micron filtration is compared with the
amount of the total rapamycin in the composition (such as a
pharmaceutical composition) prior to filtration. In some
embodiments, the recovery is assessed following storage of the
composition (such as a pharmaceutical composition) at elevated
temperatures, for example, about 40.degree. C. for about 24
hours.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) has a rapamycin recovery of at
least about any of 80%, 85%, 90%, 95%, 98%, 99%, 99.5% or more
following a 0.2 micron filtration (for example immediately after
reconstitution). In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
composition (such as a pharmaceutical composition) has a rapamycin
recovery of about any of 80-85%, 85-90%, 90-95%, 95-98%, 80-90%,
90-98%, 85-95%, 85-98%, 80-95%, 80-98%, 98%-99%, 99%-99.5%, or
99.5%-100% following a 0.2 micron filtration (for example
immediately after reconstitution).
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) has a rapamycin recovery of at
least about any of 80%, 85%, 90%, 95%, 98%, 99%, 99.5%, or more
following a 0.2 micron filtration after storage for at least about
any of 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 72 or more
hours (for example under storage at room temperature, under
refrigerated conditions, or under accelerated storage condition
(for example at about 40.degree. C.)). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the composition (such as a pharmaceutical
composition) has a rapamycin recovery of about any of 80-85%,
85-90%, 90-95%, 95-98%, 80-90%, 90-98%, 85-95%, 85-98%, 80-95%,
80-98%, 98%-99%, 99%-99.5%, or 99.5%-100% following a 0.2 micron
filtration after storage for at least about any of 6 hours, 12
hours, 24 hours, 36 hours, 48 hours, 72 hours or more hours (for
example under storage at room temperature, under refrigerated
conditions, or under accelerated storage condition (for example at
about 40.degree. C.)).
Determination of Particle Solubility
The methods of the present application in some embodiments comprise
determining particle solubility of the albumin-based nanoparticle
composition. In some embodiments, the solubility is determined
immediately after reconstitution. In some embodiments, the
solubility is determined upon storage, for example after storage
for at least about any of 6 hours, 12 hours, 24 hours, 36 hours, 48
hours, 72 hours or more hours (for example under storage at room
temperature, under refrigerated conditions, or under accelerated
storage condition (for example at 40.degree. C.)).
In some embodiments, the solubility of the pharmaceutical
composition is determined by performing dynamic light scattering
measurements for a series of concentrations of the composition
(such as a pharmaceutical composition). The proportion of intact
particles to free rapamycin is a function of the solubility of the
particles. Thus, as measured by this method, the solubility is
determined as the concentration below which particles are no longer
detectable by dynamic light scattering. In some embodiments, the
solubility is determined in a 5% human serum albumin solution. In
some embodiments, the solubility is determined in saline. In some
embodiments, the solubility is determined in plasma.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) has a solubility of about 50
.mu.g/ml to about 80 .mu.g/ml when reconstituted in a 5% human
albumin solution (for example immediately after reconstitution). In
some embodiments, the composition (such as a pharmaceutical
composition) has a solubility of about any of 40 .mu.g/ml, 45
.mu.g/ml, 50 .mu.g/ml, 55 .mu.g/ml, 60 .mu.g/ml, 65 .mu.g/ml, 70
.mu.g/ml, 75 .mu.g/ml, 80 .mu.g/ml, 85 .mu.g/ml, 90 .mu.g/ml, or
100 .mu.g/ml when reconstituted in a 5% human albumin solution (for
example immediately after reconstitution). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the composition (such as a pharmaceutical
composition) has a solubility of about any of 40-45 .mu.g/ml, 45-50
.mu.g/ml, 50-55 .mu.g/ml, 55-60 .mu.g/ml, 60-65 .mu.g/ml, 65-70
.mu.g/ml, 70-75 .mu.g/ml, 75-80 .mu.g/ml, 80-85 .mu.g/ml, 85-90
.mu.g/ml, 40-50 .mu.g/ml, 50-60 .mu.g/ml, 60-70 .mu.g/ml, 70-80
.mu.g/ml, 80-90 .mu.g/ml, 45-55 .mu.g/ml, 55-65 .mu.g/ml, 65-75
.mu.g/ml, 40-55 .mu.g/ml, 55-70 .mu.g/ml, 65-80 .mu.g/ml, 50-70
.mu.g/ml, 60-80 .mu.g/ml, 50-80 .mu.g/ml, 65-90 .mu.g/ml, 65-85
.mu.g/ml, 65-95 .mu.g/ml, 65-100 .mu.g/ml, 70-85 .mu.g/ml, 70-90
.mu.g/ml, 75-95 .mu.g/ml, or 75-100 .mu.g/ml when reconstituted in
a 5% human albumin solution (for example immediately after
reconstitution).
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) has a solubility of about 50
.mu.g/ml to about 100 .mu.g/ml when reconstituted in a 5% human
albumin solution after storage for at least about any of 6 hours,
12 hours, 24 hours, 36 hours, 48 hours, 72 hours or more hours (for
example under storage at room temperature, under refrigerated
conditions, or under accelerated storage condition (for example at
about 40.degree. C.)). In some embodiments, the composition (such
as a pharmaceutical composition) has a solubility of about any of
40 .mu.g/ml, 45 .mu.g/ml, 50 .mu.g/ml, 55 .mu.g/ml, 60 .mu.g/ml, 65
.mu.g/ml, 70 .mu.g/ml, 75 .mu.g/ml, 80 .mu.g/ml, 85 .mu.g/ml, 90
.mu.g/ml, or 100 .mu.g/ml when reconstituted in a 5% human albumin
solution after storage for at least about any of 6 hours, 12 hours,
24 hours, 36 hours, 48 hours, 72 hours or more hours (for example
under storage at room temperature, under refrigerated conditions,
or under accelerated storage condition (for example at about
40.degree. C.)). In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
composition (such as a pharmaceutical composition) has a solubility
of about any of 40-45 .mu.g/ml, 45-50 .mu.g/ml, 50-55 .mu.g/ml,
55-60 .mu.g/ml, 60-65 .mu.g/ml, 65-70 .mu.g/ml, 70-75 .mu.g/ml,
75-80 .mu.g/ml, 80-85 .mu.g/ml, 85-90 .mu.g/ml, 40-50 .mu.g/ml,
50-60 .mu.g/ml, 60-70 .mu.g/ml, 70-80 .mu.g/ml, 80-90 .mu.g/ml,
45-55 .mu.g/ml, 55-65 .mu.g/ml, 65-75 .mu.g/ml, 40-55 .mu.g/ml,
55-70 .mu.g/ml, 65-80 .mu.g/ml, 50-70 .mu.g/ml, 60-80 .mu.g/ml,
50-80 .mu.g/ml, 65-90 .mu.g/ml, 65-85 .mu.g/ml, 65-95 .mu.g/ml,
65-100 .mu.g/ml, 70-85 .mu.g/ml, 70-90 .mu.g/ml, 75-95 .mu.g/ml, or
75-100 .mu.g/ml when diluted in a 5% human albumin solution after
storage for at least about any of 6 hours, 12 hours, 24 hours, 36
hours, 48 hours, 72 hours or more hours (for example under storage
at room temperature, under refrigerated conditions, or under
accelerated storage condition (for example at about 40.degree.
C.)).
Determination of Albumin Binding Affinity
The methods of the present application in some embodiments comprise
determining albumin binding affinity to poorly water soluble drug
(such as rapamycin). In some embodiments, the binding affinity is
determined by an equilibrium dialysis test. In some embodiments,
the binding affinity is determined by Fourier transform infrared
spectroscopy (FTIR) and/or nuclear magnetic resonance (NMR)
analysis. In some embodiments, the binding affinity is determined
by an equilibrium dialysis test and FTIR. In some embodiments, the
binding affinity is determined by an equilibrium dialysis test and
NMR. In some embodiments, the binding affinity is determined by an
equilibrium dialysis test, FTIR, and NMR. In some embodiments, the
albumin binding affinity to rapamycin is determined upon
reconstitution of the pharmaceutical composition. In some
embodiments, the albumin binding affinity to rapamycin is
determined upon storage of the pharmaceutical composition.
In some embodiments, the albumin binding affinity to rapamycin is
determined by an equilibrium dialysis test. For example, the
composition (such as a pharmaceutical composition) is reconstituted
with 0.9% saline solution to create solutions containing 1 mg/mL,
75 .mu.g/mL and 50 .mu.g/mL rapamycin. Additionally, a 10% albumin
solution is prepared by diluting Albumin (Human) USP with 0.9%
sodium chloride solution. 200 .mu.L samples, with replicates, are
prepared at various rapamycin concentrations using an equilibrium
dialysis plate with 12 kD molecular weight cutoff insert. In some
embodiments, replicate wells of buffer were also included.
Subsequently, the plate is sealed and processed on an orbital
shaker, heated to 37.degree. C., for 2 hours at 800 rpm. Then, 50
.mu.L is transferred from each sample and buffer well to an empty
well on a 386-well plate for the rapamycin assay. Matrix matching
was performed by adding 50 .mu.L of 0.9% sodium chloride solution
to the wells containing samples on the 386-well plate, and by
adding 50 .mu.L of albumin solution to the wells containing buffer.
100 .mu.L of an internal rapamycin standard is added to all wells
and the plate is then processed on a plate shaker. Contents of the
plate are then transferred to a filter plate and vacuum filtered.
Subsequently, the filtrate was assayed to determine rapamycin
concentration using reversed-phase liquid chromatography mass
spectrometry. Briefly, 10 .mu.L of the filtrate was injected from
an autosampler onto a reversed-phase C18 column. Elution is
performed using a gradient method at a flow rate of 0.5 mL/min.
Mobile phase compositions are as follows: A, water with 0.1% formic
acid and B, acetonitrile with 0.1% formic acid. Effluent is
introduced into MS system through heated electrospray
ionization.
In some embodiments, the albumin binding affinity to rapamycin is
determined by an equilibrium dialysis test after dialysis for at
least 12 hours, 24 hours, 36 hours, 48 hours, 72 hours, or more
hours.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the binding affinity of
the albumin in the composition (such as a pharmaceutical
composition) for rapamycin is about the same as the binding
affinity of albumin in ABI009. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if the equilibrium binding (or dissociation) constant of the
albumin in the composition (such as a pharmaceutical composition)
for paclitaxel is less than about any of 130 .mu.M, 110 .mu.M, 90
.mu.M, 70 .mu.M, 60 .mu.M, 55 .mu.M, 50 .mu.M, or 45 .mu.M. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the binding affinity of the albumin
in the composition (such as a pharmaceutical composition) for
paclitaxel is about 42 .mu.M.
Determination of In Vitro Release Kinetics
The methods of the present application in some embodiments further
comprise determining the in vitro release kinetics of the
albumin-based nanoparticle composition. In some embodiments, the
determination is carried out immediately after reconstitution. In
some embodiments, the determination is carried out upon storage,
for example upon storage for least about any of 6 hours, 12 hours,
24 hours, 36 hours, 48 hours, 72 hours, or more hours (for example
under storage at room temperature, under refrigerated conditions,
or under accelerated storage condition (for example at about
40.degree. C.)). In some embodiments, the determination is carried
out upon storage, for example upon storage for least about any of 1
month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months,
8 months, 9 months, or more months (for example under storage at
room temperature, under refrigerated conditions, or under
accelerated storage condition (for example at about 40.degree.
C.)). In some embodiments, the determination is carried out
immediately after dilution of the composition. In some embodiments,
the determination is carried out after, for example, at least 1
hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, or more hours
after dilution of the composition.
In vitro release kinetics may be assayed using different
techniques. In one example, an in vitro release kinetics assay
measures the particle size and intensity of light scattered by the
particles using, for example, dynamic light scattering over a
period of time immediately following a reduction in particle
concentration. In some embodiments, the release kinetics are
determined by diluting the composition (such as a pharmaceutical
composition) in a 0.9% saline solution. In some embodiments, the
release kinetics is determined by diluting the composition (such as
a pharmaceutical composition) in a 5% human serum albumin solution.
In a second example, an in vitro release kinetics assay measures
the absorbance of the composition over a period of time immediately
following a reduction in particle concentration. In some
embodiments, the absorbance of the composition is measured using a
UV-Vis spectrophotometer. In some embodiments, the absorbance of
the composition is measured using a UV-Vis spectrophotometer
equipped with a 295 nm cut-off filter. In some embodiments, the
absorbance of the composition is measured at, for example, 340 nm.
In some embodiments, the composition is diluted to, for example,
100 .mu.g/ml, as measured by the concentration of rapamycin.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the dissolution profile
of the composition (such as a pharmaceutical composition), as
measured by an in vitro release kinetics assay, matches the
dissolution profile of ABI-009. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the dissolution profile of the composition (such as
a pharmaceutical composition) following an accelerated aging
process matches the dissolution profile of ABI-009. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the dissolution profile of the
composition (such as a pharmaceutical composition) following an
accelerated aging process matches the dissolution profile of
ABI-009 following the same accelerated aging process. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the composition (such as a
pharmaceutical composition) has an in vitro release kinetic
behavior that is similar to that of ABI-009 under the same assay
conditions upon storage for at least about any of 6 hours, 12
hours, 24 hours, 36 hours, 48 hours, 72 hours, or more hours (for
example under storage at room temperature, under refrigerated
conditions, or under accelerated storage condition (for example at
about 40.degree. C.)).
In some embodiments, the in vitro release is determined using the
dynamic light scattering method. In some embodiments, the in vitro
release kinetics assay measures the intensity of light scattered by
the composition over a period of time immediately following a
reduction in particle concentration. In some embodiments, the light
scattering intensity of the composition is measured using a dynamic
light scattering apparatus, where the concentration of rapamycin
released from the nanoparticles is calculated from the intensity of
scattered light. In some embodiments, the intensity of light
scattered is measured at a scattering angle of 173.degree.. In some
embodiments, the composition is diluted to 20 .mu.g/ml, as measured
by the concentration of rapamycin, and in some embodiments the
composition is diluted to 37.5 .mu.g/ml. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the mean value of percent released rapamycin from a
nanoparticle composition (such as a pharmaceutical composition) at
20 .mu.g/ml in 10 mM sodium chloride, as measured by the rapamycin
concentration, after 120 minutes is about 100%. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the mean value of percent released
rapamycin from a nanoparticle composition (such as a pharmaceutical
composition) at 37.5 .mu.g/ml in 10 mM sodium chloride, as measured
by the rapamycin concentration, after 120 minutes is about 85%.
Determination of Physical Stability
The methods of the present application in some embodiments comprise
determining the physical stability of particles in the composition
(such as a pharmaceutical composition). In some embodiments, the
stability is determined immediately after reconstitution. In some
embodiments, the stability is measured upon storage, for example
upon storage for at least about any of 6 hours, 12 hours, 24 hours,
36 hours, 48 hours, 72 hours, or more hours (for example under
storage at room temperature, under refrigerated conditions, or
under accelerated storage condition (for example at about
40.degree. C.)). In some embodiments, the stability is measured
after storage, for example, for 8 hours at about 5.degree. C.
followed by storage for 8 hours at about 25.degree. C., or after
for storage for 24 hours at about 25.degree. C.
Stability of nanoparticles in the pharmaceutical composition can be
assayed by a number of techniques, including, but not limited to,
visual inspection (such as visual appearance, visual color, visible
particulate matter), microscopy imaging, and loss of potency. In
some embodiments, the stability of the composition (such as a
pharmaceutical composition) is determined based on sedimentation.
Sedimentation can be assessed by visual inspection and/or
microscopy (such as cross-polarization microscopy). Microscopy can
be used to determine the size of aggregated sediment particles. In
some embodiments, the stability of the composition (such as a
pharmaceutical composition) is determined based on the crystalline
state of the nanoparticles, for example by increased presence of
nanoparticles with crystalline rapamycin. In some embodiments, the
stability of the composition (such as a pharmaceutical composition)
is determined based on a loss of potency following a 0.2 micron
filtration of the composition (such as a pharmaceutical
composition). In some embodiments, the loss of potency is in vitro
potency. In some embodiments, the loss of potency is in vivo
potency.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the nanoparticles are
physically stable (for example immediately after reconstitution).
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the nanoparticles are
physically stable for at least about any of 6 hours, 12 hours, 24
hours, 36 hours, 48 hours, 72 hours or more hours upon storage (for
example under storage at room temperature, under refrigerated
conditions, or under accelerated storage condition (for example at
about 40.degree. C.)). In some embodiments, the composition (such
as a pharmaceutical composition) is suitable for medical use if the
nanoparticles are physically stable after storage for 8 hours at
about 5.degree. C. followed by storage for 8 hours at about
25.degree. C., or after for storage for 24 hours at about
25.degree. C.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) shows no visible particulate
matter (for example immediately after reconstitution). In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the composition (such as a
pharmaceutical composition) shows no visible particulate matter for
at least about any of 6 hours, 12 hours, 24 hours, 36 hours, 48
hours, 72 hours or more hours upon storage (for example under
storage at room temperature, under refrigerated conditions, or
under accelerated storage condition (for example at about
40.degree. C.)). In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
composition (such as a pharmaceutical composition) shows no visible
particulate matter after storage for 8 hours at about 5.degree. C.
followed by storage for 8 hours at about 25.degree. C., or after
for storage for 24 hours at about 25.degree. C.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) shows no sedimentation (for
example immediately after reconstitution). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the composition (such as a pharmaceutical
composition) shows no sedimentation for at least about any of 6
hours, 12 hours, 24 hours, 36 hours, 48 hours, 72 hours or more
hours upon storage (for example under storage at room temperature,
under refrigerated conditions, or under accelerated storage
condition (for example at about 40.degree. C.)). In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for medical use if the composition (such as a
pharmaceutical composition) shows no sedimentation after storage
for 8 hours at about 5.degree. C. followed by storage for 8 hours
at about 25.degree. C., or after for storage for 24 hours at about
25.degree. C.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the rapamycin in the
composition (such as a pharmaceutical composition) shows no
crystallinity (e.g., by polarized light microscopy) (for example
immediately after reconstitution). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the rapamycin in the composition (such as a
pharmaceutical composition) shows no crystallinity (e.g., by
polarized light microscopy) for at least about any of 6 hours, 12
hours, 24 hours, 36 hours, 48 hours, 72 hours or more hours upon
storage (for example under storage at room temperature, under
refrigerated conditions, or under accelerated storage condition
(for example at about 40.degree. C.)). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the rapamycin in the composition (such as a
pharmaceutical composition) shows no crystallinity (e.g., by
polarized light microscopy) after storage for 8 hours at about
5.degree. C. followed by storage for 8 hours at about 25.degree.
C., or after for storage for 24 hours at about 25.degree. C.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) shows no loss of potency (e.g., by
in vitro or in vivo testing) following a 0.2 micron filtration (for
example immediately after reconstitution). In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the composition (such as a pharmaceutical
composition) shows no loss of potency (e.g., by in vitro or in vivo
testing) for at least about any of 6 hours, 12 hours, 24 hours, 36
hours, 48 hours, 72 hours or more hours upon storage (for example
under storage at room temperature, under refrigerated conditions,
or under accelerated storage condition (for example at about
40.degree. C.)). In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if the
composition (such as a pharmaceutical composition) shows no loss of
potency (e.g., by in vitro or in vivo testing) after storage for 8
hours at about 5.degree. C. followed by storage for 8 hours at
about 25.degree. C., or after for storage for 24 hours at about
25.degree. C.
Determination of Osmolality
The methods described herein in some embodiments comprise
determining the osmolality of a reconstituted composition (such as
a pharmaceutical composition). In some embodiments, the osmolality
is determined immediately after reconstitution. In some
embodiments, the osmolality is measured upon storage, for example
upon storage for at least about any of 1 month, 2 months, 3 months,
4 months, 5 months, or 6 months, at a temperature of, for example,
about 25.degree. C. or about 40.degree. C. In some embodiments, the
osmolality is measured after storage for 6 months at about
25.degree. C. and a relative humidity of about 60% at an inverted
position. In some embodiments, the osmolality is measured after
storage for 6 months at about 40.degree. C. and a relative humidity
of about 75% at an inverted position.
Osmolality of a composition (such as a pharmaceutical composition)
can be assayed by a number of methods, including, comprising use of
a vapor pressure depression osmometer, a membrane osmometer, or a
freezing point depression osmometer.
In some embodiments, the composition (such as a pharmaceutical
composition) has an osmolality of between about 320 mOsm/kg to
about 360 mOsm/kg. In some embodiments, the composition (such as a
pharmaceutical composition) has an osmolality of between about 325
mOsm/kg to about 355 mOsm/kg. In some embodiments, the composition
(such as a pharmaceutical composition) has an osmolality of between
about 330 mOsm/kg to about 350 mOsm/kg. In some embodiments, the
composition (such as a pharmaceutical composition) has an
osmolality of about 330 mOsm/kg, 331 mOsm/kg, 332 mOsm/kg, 333
mOsm/kg, 334 mOsm/kg, 335 mOsm/kg, 336 mOsm/kg, 337 mOsm/kg, 338
mOsm/kg, 339 mOsm/kg, 340 mOsm/kg, 341 mOsm/kg, 342 mOsm/kg, 343
mOsm/kg, 344 mOsm/kg, 345 mOsm/kg, 346 mOsm/kg, 347 mOsm/kg or 348
mOsm/kg.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) shows no significant change in
osmolality after storage. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if the composition (such as a pharmaceutical composition) shows no
significant change in osmolality after storage at elevated
temperatures, such as 40.degree. C. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the composition (such as a pharmaceutical
composition) shows no significant change in osmolality for at least
about any of 1 month, 2 months, 3 months, 4 months, 5 months, 6
months or more months upon storage (for example under storage at
room temperature, under refrigerated conditions, or under
accelerated storage condition (for example at about 40.degree.
C.)).
Determination of Viscosity
The methods described herein in some embodiments comprise
determining the viscosity (such as dynamic viscosity) of a
reconstituted composition (such as a pharmaceutical composition).
In some embodiments, the viscosity is determined immediately after
reconstitution. In some embodiments, the viscosity is measured upon
storage, for example upon storage for at least about any of 1
month, 2 months, 3 months, 4 months, 5 months, or 6 months, at a
temperature of, for example, about 25.degree. C. or about
40.degree. C. In some embodiments, the viscosity is measured after
storage for 6 months at about 25.degree. C. and a relative humidity
of about 60% at an inverted position. In some embodiments, the
viscosity is measured after storage for 6 months at about
40.degree. C. and a relative humidity of about 75% at an inverted
position.
Viscosity (such as dynamic viscosity) of a composition (such as a
pharmaceutical composition) can be assayed by a number of methods,
including, comprising use of a viscometers and rheometers.
In some embodiments, the composition (such as a pharmaceutical
composition) has a viscosity of between about 1.20 centipoise to
about 1.50 centipoise. In some embodiments, the composition (such
as a pharmaceutical composition) has a viscosity of between about
1.25 centipoise to about 1.45 centipoise. In some embodiments, the
composition (such as a pharmaceutical composition) has a viscosity
of about 1.25 centipoise, 1.26 centipoise, 1.27 centipoise, 1.28
centipoise, 1.29 centipoise, 1.30 centipoise, 1.31 centipoise, 1.32
centipoise, 1.33 centipoise, 1.34 centipoise, 1.35 centipoise, 1.36
centipoise, 1.37 centipoise, 1.38 centipoise, 1.39 centipoise, 1.40
centipoise, 1.41 centipoise, 1.42 centipoise, 1.43 centipoise, 1.44
centipoise, or 1.45 centipoise.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if the composition (such
as a pharmaceutical composition) shows no significant change in
viscosity after storage. In some embodiments, the composition (such
as a pharmaceutical composition) is suitable for medical use if the
composition (such as a pharmaceutical composition) shows no
significant change in viscosity after storage at elevated
temperatures, such as 40.degree. C. In some embodiments, the
composition (such as a pharmaceutical composition) is suitable for
medical use if the composition (such as a pharmaceutical
composition) shows no significant change in viscosity for at least
about any of 1 month, 2 months, 3 months, 4 months, 5 months, 6
months or more months upon storage (for example under storage at
room temperature, under refrigerated conditions, or under
accelerated storage condition (for example at about 40.degree.
C.)).
Determination of Tumor Distribution of Rapamycin In Vivo
The methods described herein in some embodiments comprise
determining tumor distribution of poorly water soluble drug (such
as rapamycin) upon administration of the albumin-based rapamycin
nanoparticle composition in vivo. Variations in albumin-based
nanoparticle compositions, and their formulations, can affect in
vivo distribution of poorly water soluble drug (such as rapamycin)
within tumor tissue. In some embodiments, tumor penetration of a
composition (such as a pharmaceutical composition) is determined.
In some embodiments, tumor cell uptake of a composition (such as a
pharmaceutical composition) is determined. In some embodiments,
tumor penetration and tumor cell update of a composition (such as a
pharmaceutical composition) are determined.
In some embodiments, the method comprises administering the
composition (such as a pharmaceutical composition) to an animal
having a tumor and measuring the level of rapamycin in the tumor
tissue. The level of rapamycin can be measured, for example, by
imaging, immunohistochemistry or quantitative mass spectrometry
methods. In some embodiments, the rapamycin in the composition
(such as a pharmaceutical composition) that is administered into
the animal is radio-labeled, thus allowing sensitive determination
of tumor distribution of the rapamycin.
In some embodiments, methods for determining the distribution of
rapamycin within tumor tissue comprise use of a tumor model. In
some embodiments, the methods are performed on a naturally
occurring tumor in an individual. In some embodiments, the methods
are performed on a xenograft tumor in an individual. In some
embodiments, the tumor model is a MIA PaCa-2 xenograft, A2058
xenograft, or H2122 xenograft. In some embodiments, the
distribution of rapamycin within tumor tissue is determined upon
reconstitution of the pharmaceutical composition. In some
embodiments, the distribution of rapamycin within tumor tissue is
determined upon storage of the pharmaceutical composition.
In some embodiments, the composition (such as a pharmaceutical
composition) is administered to the animal by systemic
administration. In some embodiments, the composition (such as a
pharmaceutical composition) is delivered to the tumor by direct
injection. In some embodiments, the composition (such as a
pharmaceutical composition) is delivered to the tumor by direct
microinjection, for example, via an arrayed microinjection device
such as those sold under the trademark CIVO.RTM. (Presage
Biosciences, Seattle, Wash.). In some embodiments, the composition
(such as a pharmaceutical composition) is delivered into flank
tumor xenografts. For methods comprising the use of direct
injection, in some embodiments, an imaging agent is injected with
the composition (such as a pharmaceutical composition) to locate
the injection site. In some embodiments, the composition (such as a
pharmaceutical composition) is co-injected with an imaging agent,
such as a fluorescent compound, for example, a fluorochrome sold
under the trademark VIVOTAG-S.RTM. 680. In some embodiments, a
labeled-rapamycin is used in the composition (such as a
pharmaceutical composition).
In some embodiments, methods for determining the distribution of
rapamycin within tumor tissue comprise measuring spatial
distribution of rapamycin within the tumor and/or tumor-associated
tissue. In some embodiments, spatial distribution of rapamycin is
measured from sections (e.g., slices) of tumor tissue. In some
embodiments, the sections of tumor tissue are perpendicular to the
injection path. In some embodiments, spatial distribution of
rapamycin is measured by imaging, such as immunohistochemical
imaging, fluorescent imaging, or any combinations thereof, of the
tumor tissue. In some embodiments, spatial distribution of
rapamycin is measured by immunohistochemical staining of cells in
mitotic arrest. In some embodiments, cells in mitotic arrest are
detected by phospho-histone H3 (pHH3) staining, such as
immunohistochemical staining of pHH3 using an anti-pHH3 antibody.
Generally, a cell stained by a pHH3 detection agent is indicative
of a cell in mitotic arrest. In some embodiments, tumor tissue is
stained with a cell imaging agent, such as an agent for imaging a
cell nucleus, for example, 4',6-diamidino-2-phenylindole (DAPI). In
some embodiments, spatial distribution of rapamycin comprises use
of software, such as that sold under the trademark CIVOANALYZER.TM.
(Presage Biosciences, Seattle, Wash.). In some embodiments, spatial
distribution of rapamycin is measured by performing mass
spectrometry-based imaging (e.g., MALDI-MS-based imaging). In some
embodiments, spatial distribution of rapamycin is measured by
detecting labeled-rapamycin.
In some embodiments, spatial distribution of rapamycin in a tumor
tissue is performed on more than one tumor sections (e.g., slices)
from a tumor. In some embodiments, spatial distribution of
rapamycin in tumor tissue is performed on more than one adjacent
tumor sections from a tumor, thereby allowing for the recreation of
a 3-dimensional tumor distribution of rapamycin. In some
embodiments, the more than one adjacent tumor sections are in
spatial proximity to a microinjection site.
In some embodiments, distribution of rapamycin within tumor tissue
is determined by measuring spatial distribution of rapamycin over a
time-course following administration of the composition (such as a
pharmaceutical composition). In some embodiments, distribution of
rapamycin within tumor tissue is measured at about 24 hours, about
48 hours, and/or about 72 hours following administration. In some
embodiments, distribution of rapamycin within tumor tissue is
measured at least about 12 hours, at least about 24 hours, at least
about 48 hours, and/or at least about 72 hours following
administration. In some embodiments, distribution of rapamycin
within tumor tissue is determined by direct microinjection of a
pharmaceutical composition into flank human tumor xenografts and
analyzed, for example, at about 24 hours, about 48 hours, and/or
about 72 hours following direct microinjection.
In some embodiments, the distribution of rapamycin within tumor
tissue is determined based on detection of rapamycin in an area of
the tumor tissue. In some embodiments, the distribution of
rapamycin within tumor tissue is determined based on detection of
rapamycin within a cell. Generally, methods comprising
identification of cells in mitotic arrest comprise assessing the
number of cells in mitotic arrest and/or the number of cells not in
mitotic arrest. In some embodiments, the distribution of rapamycin
is determined based on the percentage of cells in mitotic arrest.
In some embodiments, the percentage of mitotic arrest is reported
as the fraction of pHH3 positive cells (i.e., number of pHH3
positive cells over the number of total cells in the sample area).
In some embodiments, mitotic arrest is measured at one or more
defined radial distances extending from the site of injection, for
example, at about 100 .mu.m, about 200 .mu.m, about 300 .mu.m,
about 400 .mu.m, about 500 .mu.m, about 600 .mu.m, about 700 .mu.m,
about 800 .mu.m, about 900 .mu.m, about 1000 .mu.m, about 1100
.mu.m, about 1200 .mu.m, and/or about 1300 .mu.m from the site of
the injection. In some embodiments, mitotic arrest is measured at
one or more defined radial distances extending from the site of
injection, for example, at least about 100 .mu.m, at least about
200 .mu.m, at least about 300 .mu.m, at least about 400 .mu.m, at
least about 500 .mu.m, at least about 600 .mu.m, at least about 700
.mu.m, at least about 800 .mu.m, at least about 900 .mu.m, at least
about 1000 .mu.m, at least about 1100 .mu.m, at least about 1200
.mu.m, and/or at least about 1300 .mu.m from the site of the
injection.
In some embodiments, the distribution of rapamycin within tumor
tissue comprises determining an area of response (e.g., an area of
tumor tissue wherein rapamycin is detected).
In some embodiments, the distribution profile of rapamycin within
tumor tissue for a composition (such as a pharmaceutical
composition) is compared to the distribution profile of rapamycin
for another composition. Generally, the amount (e.g.,
concentration) of rapamycin in each composition is measured, for
example by mass spectrometry, to allow for normalization of
rapamycin concentrations between compositions (such as
pharmaceutical compositions).
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if it allows an enhanced
rapamycin tumor distribution. A composition allows "enhanced
rapamycin tumor distribution" if, upon tumor injection, it allows a
distribution of rapamycin within the tumor tissue that is more
extensive than that of a solvent-based rapamycin (such as rapamycin
in DMSO) formulation. In some embodiments, the radial distance of
rapamycin spread (penetration and/or tumor cell uptake of
rapamycin) in a tumor tissue extending from the site of injection
of the composition (such as a pharmaceutical composition) is
greater than (for example more than about 1.1.times., 1.2.times.,
1.3.times., 1.4.times., 1.5.times., 1.6.times., 1.7.times.,
1.8.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
7.times., 8.times., or more of) that of a solvent-based rapamycin
(such as rapamycin in DMSO) formulation under the same assay
conditions.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if upon tumor injection it
allows a distribution of rapamycin within the tumor tissue that is
similar to that of ABI-009 under the same assay conditions. For
example, in some embodiments, the radial distance of rapamycin
penetration (or tumor cell uptake of rapamycin) in a tumor tissue
extending from the site of injection of the composition (such as a
pharmaceutical composition) is greater than or equal to that of
ABI-009 under the same assay conditions. In some embodiments, the
radial distance of rapamycin penetration (or tumor cell uptake of
rapamycin) in a tumor tissue extending from the site of injection
of the composition (such as a pharmaceutical composition) is at
least about 90% (including for example at least about any of 90%,
95%, 98% or 99%) of that of ABI-009 under the same assay
conditions.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if it allows rapamycin to
spread radially for more than about 700 .mu.m (such as more than
about 1200 .mu.m) within about 24 hours when the composition (such
as a pharmaceutical composition) is injected into a tumor tissue
(for example when injected at the rapamycin amount of about 12
.mu.g (or 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor).
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if it allows rapamycin to
spread radially for more than about 700 .mu.m within about any one
of 12 hours, 24 hours, 36 hours, 48 hours, or 72 hours when the
composition (such as a pharmaceutical composition) is injected into
a tumor tissue. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if it
allows rapamycin to spread radially for more than about 1200 .mu.m
within about any one of 12 hours, 24 hours, 36 hours, 48 hours, or
72 hours when the composition (such as a pharmaceutical
composition) is injected into a tumor tissue. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if it allows rapamycin to spread radially for more
than about any of 200 .mu.m, 300 .mu.m, 400 .mu.m, 500 .mu.m, 600
.mu.m, 700 .mu.m, 750 .mu.m, 800 .mu.m, 850 .mu.m, 900 .mu.m, 950
.mu.m, 1000 .mu.m, 1100 .mu.m, 1200 .mu.m, 1300 .mu.m, 1400 .mu.m,
1500 .mu.m or 2000 .mu.m within about 12 hours when the composition
(such as a pharmaceutical composition) is injected into a tumor
tissue. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if it
allows rapamycin to spread radially for more than about any of 200
.mu.m, 300 .mu.m, 400 .mu.m, 500 .mu.m, 600 .mu.m, 700 .mu.m, 750
.mu.m, 800 .mu.m, 850 .mu.m, 900 .mu.m, 950 .mu.m, 1000 .mu.m, 1100
.mu.m, 1200 .mu.m, 1300 .mu.m, 1400 .mu.m, 1500 .mu.m or 2000 .mu.m
within about 24 hours when the composition (such as a
pharmaceutical composition) is injected into a tumor tissue. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if it allows rapamycin to
spread radially for more than about any of 200 .mu.m, 300 .mu.m,
400 .mu.m, 500 .mu.m, 600 .mu.m, 700 .mu.m, 750 .mu.m, 800 .mu.m,
850 .mu.m, 900 .mu.m, 950 .mu.m, 1000 .mu.m, 1100 .mu.m, 1200
.mu.m, 1300 .mu.m, 1400 .mu.m, 1500 .mu.m or 2000 .mu.m within
about 36 hours when the composition (such as a pharmaceutical
composition) is injected into a tumor tissue. In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if it allows rapamycin to spread radially for more
than about any of 200 .mu.m, 300 .mu.m, 400 .mu.m, 500 .mu.m, 600
.mu.m, 700 .mu.m, 750 .mu.m, 800 .mu.m, 850 .mu.m, 900 .mu.m, 950
.mu.m, 1000 .mu.m, 1100 .mu.m, 1200 .mu.m, 1300 .mu.m, 1400 .mu.m,
1500 .mu.m or 2000 .mu.m within about 48 hours when the composition
(such as a pharmaceutical composition) is injected into a tumor
tissue. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if it
allows rapamycin to spread radially for more than about any of 200
.mu.m, 300 .mu.m, 400 .mu.m, 500 .mu.m, 600 .mu.m, 700 .mu.m, 750
.mu.m, 800 .mu.m, 850 .mu.m, 900 .mu.m, 950 .mu.m, 1000 .mu.m, 1100
.mu.m, 1200 .mu.m, 1300 .mu.m, 1400 .mu.m, 1500 .mu.m or 2000 .mu.m
within about 72 hours when the composition (such as a
pharmaceutical composition) is injected into a tumor tissue. In
some embodiments, the composition (such as a pharmaceutical
composition) meets two or more of the above-described functional
attributes.
Determination of the radial distance of rapamycin penetration (or
tumor cell uptake) extending from the injection site in a tumor
tissue requires setting a baseline percentage (or fraction) of
mitotically arrested cells among all cells in the tumor tissue. The
radial distance of rapamycin penetration (or tumor cell uptake)
extending from the injection site is the radial distance extending
from the injection site, beyond which the percentage of mitotically
arrested cells among all cells at the radial distance is no more
than the baseline percentage. In some embodiments, the baseline
percentage is set to be about 0.6%. In some embodiments, the
baseline percentage is set to be about any of 0.1%, 0.3%, 0.5%,
0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 5%, 10%,
or more. The radial distance of rapamycin penetration (or tumor
cell uptake) extending from the injection site of the composition
(such as a pharmaceutical composition), and that of ABI-009 are
compared to determine whether tumor distribution of the composition
(such as a pharmaceutical composition) in vivo is similar to that
of ABI-009. Similarly, the radial distance of rapamycin penetration
(or tumor cell uptake) extending from the injection site of the
composition (such as a pharmaceutical composition), and that of a
solvent-based rapamycin (such as rapamycin in DMSO) are compared to
determine whether tumor distribution of the composition (such as a
pharmaceutical composition) in vivo is greater than that of a
solvent-based rapamycin (such as rapamycin in DMSO).
In some embodiments, tumor distribution of rapamycin is determined
based on the percentage of mitotically arrested cells (such as
pHH3+ cells) at a defined radial distance extending from the site
of injection. The percentages (or fractions) of mitotically
arrested cells (such as pHH3+ cells) at defined radial distances
extending from the site of injection of the compositions (such as
pharmaceutical compositions) vary according to the volume and dose
of the rapamycin in the composition (such as a pharmaceutical
composition) being injected, the time at which the tumor tissue is
harvested, the cell type of the tumor tissue or xenograft (such as
cell lines or source of tumor tissue), the animal model, or other
assay conditions.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if, upon injection of the
pharmaceutical composition, the percentage (or fraction) of
mitotically arrested cells (such as pHH3+ cells) at a radial
distance extending from the site of injection (such as at any of
200 .mu.m, 400 .mu.m, 500 .mu.m, 600 .mu.m, 700 .mu.m, 800 .mu.m,
900 .mu.m, 1000 .mu.m, 1100 .mu.m, 1200 .mu.m, 1500 .mu.m, or 2000
.mu.m) of the composition (such as a pharmaceutical composition) is
similar to that of ABI-009 within about 24 hours of injection. In
some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if, upon injection of the
pharmaceutical composition, the percentages (or fractions) of
mitotically arrested cells (such as pHH3+ cells) at a plurality of
radial distances extending from the site of injection (such as at
any combination of 200 .mu.m, 400 .mu.m, 500 .mu.m, 600 .mu.m, 700
.mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m, 1200 .mu.m,
1500 .mu.m, or 2000 .mu.m) of the composition (such as a
pharmaceutical composition) is similar to those of ABI-009 within
about 24 hours of injection. In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if, upon injection of the pharmaceutical composition, the
percentage (or fraction) of mitotically arrested cells (such as
pHH3+ cells) at a radial distance extending from the site of
injection (such as at any of 200 .mu.m, 400 .mu.m, 500 .mu.m, 600
.mu.m, 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m,
1200 .mu.m, 1500 .mu.m, or 2000 .mu.m) of the composition (such as
a pharmaceutical composition) is greater than that of a
solvent-based rapamycin (such as rapamycin in DMSO) within about 24
hours of injection. In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if, upon
injection of the pharmaceutical composition, the percentages (or
fractions) of mitotically arrested cells (such as pHH3+ cells) at a
plurality of radial distances extending from the site of injection
(such as at any combination of 200 .mu.m, 400 .mu.m, 500 .mu.m, 600
.mu.m, 700 .mu.m, 800 .mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m,
1200 .mu.m, 1500 .mu.m, or 2000 .mu.m) of the composition (such as
a pharmaceutical composition) are greater than those of a
solvent-based rapamycin (such as rapamycin in DMSO) within about 24
hours of injection.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if it allows more than
about 5% (such as more than about any of 4%, 4.5%, 5%, 5.5%, 6%,
6.5%, or 7% or more) of all cells at a radial distance of about 400
.mu.m extending from a site of injection to be mitotically arrested
cells within about 24 hours of the injection when the composition
(such as a pharmaceutical composition) is injected into a tumor
tissue (for example when injected at the rapamycin amount of about
12 .mu.g (or 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft
tumor). In some embodiments, the composition (such as a
pharmaceutical composition) is suitable for medical use if it
allows more than about 3.5% (such as more than about any one of 2%,
2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5% or more) of all cells at a
radial distance of about 400 .mu.m extending from a site of
injection to be mitotically arrested cells within about 24 hours of
the injection when the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example when
injected at the rapamycin amount of about 12 .mu.g (or 4 mg/ml)
into a pancreatic MIA PaCa-2 xenograft tumor). In some embodiments,
the composition (such as a pharmaceutical composition) is suitable
for medical use if it allows more than about 2.5% (such as more
than about any one of 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5% or
more) of all cells at a radial distance of about 400 .mu.m
extending from a site of injection to be mitotically arrested cells
within about 24 hours of the injection when the composition (such
as a pharmaceutical composition) is injected into a tumor tissue
(for example when injected at the rapamycin amount of about 12
.mu.g (or 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor).
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for medical use if it allows more than
about 1.5% (such as more than about any one of 0.5%, 0.8%, 1%,
1.5%, 2%, 2.5%, 3% or more) of all cells at a radial distance of
about 400 .mu.m extending from a site of injection to be
mitotically arrested cells within about 24 hours of the injection
when the composition (such as a pharmaceutical composition) is
injected into a tumor tissue (for example when injected at the
rapamycin amount of about 12 .mu.g (or 4 mg/ml) into a pancreatic
MIA PaCa-2 xenograft tumor). In some embodiments, the composition
(such as a pharmaceutical composition) is suitable for medical use
if it allows more than about 1% (such as more than about any one of
0.4%, 0.6%, 0.8%, 1%, 1.5%, 2% or more) of all cells at a radial
distance of about 400 .mu.m extending from a site of injection to
be mitotically arrested cells within about 24 hours of the
injection when the composition (such as a pharmaceutical
composition) is injected into a tumor tissue (for example when
injected at the rapamycin amount of about 12 .mu.g (or 4 mg/ml)
into a pancreatic MIA PaCa-2 xenograft tumor). In some embodiments,
suitability of medical use is determined based on two or more of
the above-described functional attributes.
Thus, the present application in one aspect also provides methods
of determining distribution of rapamycin in a tumor tissue. For
example, in some embodiments there is provided a method of
determining distribution of rapamycin in a tumor tissue upon
administration of an albumin-based rapamycin nanoparticle
composition to the tumor tissue, comprising 1) injecting the
composition (such as a pharmaceutical composition) directly to the
tumor tissue; and 2) assessing the spatial distribution of
rapamycin within the tumor tissue. In some embodiments, there is
provided a method of determining distribution of rapamycin in a
tumor tissue upon administration of an albumin-based rapamycin
nanoparticle composition to the tumor tissue, comprising 1)
microinjecting (for example by using an arrayed microinjection
device such as those sold under the trademark CIVO.RTM. (Presage
Biosciences, Seattle, Wash.)) the composition (such as a
pharmaceutical composition) directly to the tumor tissue; and 2)
assessing the spatial distribution of rapamycin within the tumor
tissue. In some further embodiments, the injection further
comprises injection of an imaging agent. In some embodiments, the
composition (such as a pharmaceutical composition) is co-injected
with an imaging agent to determine the location of the injection
site. In some embodiments, the composition (such as a
pharmaceutical composition) is co-injected with a fluorochrome sold
under the trademark VIVOTAG-S.RTM. 680 to determine the location of
the injection site.
In some embodiments, there is provided a method of determining
distribution of rapamycin in a tumor tissue upon administration of
an albumin-based rapamycin nanoparticle composition to the tumor
tissue, comprising 1) microinjecting (for example by using an
arrayed microinjection device such as those sold under the
trademark CIVO.RTM. (Presage Biosciences, Seattle, Wash.) the
composition (such as a pharmaceutical composition) directly to the
tumor tissue; and 2) staining the tissue with an agent that
indicate the presence of rapamycin; and 3) determining the radial
distance of rapamycin distribution extending from the site of
injection. In some embodiments, the agent that indicates the
presence of rapamycin is an indicator of mitotic arrest such as an
anti-pHH3 detection agent (for example an antibody).
In some embodiments, there is provided a method of determining
distribution of rapamycin in a tumor tissue upon administration of
an albumin-based rapamycin nanoparticle composition to the tumor
tissue, comprising 1) microinjecting (for example by using an
arrayed microinjection device such as those sold under the
trademark CIVO.RTM. (Presage Biosciences, Seattle, Wash.) the
composition (such as a pharmaceutical composition) directly to the
tumor tissue; and 2) staining the tissue with an agent that
indicate mitotic arrest of a cell; and 3) determining the radial
distance of rapamycin distribution extending from the site of
injection.
In some embodiments, there is provided a method of determining
distribution of rapamycin in a tumor tissue upon administration of
an albumin-based rapamycin nanoparticle composition to the tumor
tissue, comprising 1) microinjecting (for example by using an
arrayed microinjection device such as those sold under the
trademark CIVO.RTM. (Presage Biosciences, Seattle, Wash.) the
composition (such as a pharmaceutical composition) directly to the
tumor tissue; and 2) staining the tissue with an agent that
indicate the mitotic arrest of the cell; and 3) determining the
fraction of stained cells a given radial distance extending from
the injection site. In some embodiments, the agent that indicates
mitotic arrest of a cell is an anti-pHH3 detection agent (for
example an antibody).
In some embodiments, there is provided a method of determining
distribution of rapamycin in a tumor tissue upon administration of
an albumin-based rapamycin nanoparticle composition to the tumor
tissue, comprising 1) microinjecting (for example by using an
arrayed microinjection device such as those sold under the
trademark CIVO.RTM. (Presage Biosciences, Seattle, Wash.) the
composition (such as a pharmaceutical composition) directly to the
tumor tissue; and 2) staining the tissue with an agent that
indicate the mitotic arrest of the cell; and 3) determining the
fraction of cells in mitotic arrest at a given radial distance
extending from the injection site. In some embodiments, the agent
that indicates mitotic arrest of a cell is an anti-pHH3 detection
agent (for example an antibody).
The albumin-based nanoparticle compositions assessed by methods
descried herein in some embodiments have a specific albumin to
poorly water soluble drug (such as rapamycin) ratio. For example,
in some embodiments, the weight ratio of the total albumin to the
total rapamycin in the composition (such as a pharmaceutical
composition) is about 3:1 to about 7.9:1 or about 10:1 to about
17:1. In some embodiments, the weight ratio of the total albumin to
the total rapamycin in the composition (such as a pharmaceutical
composition) is about 3:1 to about 7.9:1, which including for
example about 4:1 to about 7:1, about 5:1 to about 7:1, about 6:1
to about 7:1, about 7:1 to about 7.5:1, and about 7.5:1 to about
7.9:1. In some embodiments, the weight ratio of the total albumin
to the total rapamycin in the composition (such as a pharmaceutical
composition) is about 10:1 to about 17:1, which include for
example, about 10:1 to about 15:1, about 10:1 to about 12:1, about
10:1 to about 11:1, or about 10:1 to about 10.5:1.
In some embodiments, the weight ratio of the total albumin to the
total rapamycin in the composition (such as a pharmaceutical
composition) is about 8:1 to about 10:1. In some embodiments, the
weight ratio of the total albumin to the total rapamycin in the
composition (such as a pharmaceutical composition) is about
9:1.
In some embodiments, the weight ratio of the total albumin to the
total rapamycin in the composition (such as a pharmaceutical
composition) is about any of 1:1, 2:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1,
5.5:1, 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, 9:1, 9.5:1, 10:1,
10.5:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or
20:1. In some embodiments, the weight ratio of the total albumin to
the total rapamycin in the composition (such as a pharmaceutical
composition) is any of about 1:1 to about 2:1, about 2:1 to about
3:1, about 3:1 to about 3.5:1, about 3.5:1 to about 4:1, about 4:1
to about 4.5:1, about 4.5:1 to about 5:1, about 5:1 to about 5.5:1,
about 5.5:1 to about 6:1, about 6:1 to about 6.5:1, about 6.5:1 to
about 7:1, about 7:1 to about 7.5:1, about 7.5:1 to about 8:1,
about 8:1 to about 8.5:1, about 8.5:1 to about 9:1, about 9:1 to
about 9.5:1, about 9.5:1 to about 10:1, about 10:1 to about 10.5:1,
about 10.5:1 to about 11:1, about 11:1 to about 12:1, about 12:1 to
about 13:1, about 13:1 to about 14:1, about 14:1 to about 15:1,
about 15:1 to about 16:1, about 16:1 to about 17:1, about 17:1 to
about 18:1, about 18:1 to about 19:1, about 19:1 to about 20:1,
about 1:1 to about 3:1, about 3:1 to about 4:1, about 4:1 to about
5:1, about 5:1 to about 6:1, about 6:1 to about 7:1, about 7:1 to
about 8:1, about 8:1 to about 9:1, about 9:1 to about 10:1, about
10:1 to about 11:1, about 11:1 to about 13:1, about 13:1 to about
15:1, about 15:1 to about 17:1, about 17:1 to about 19:1, about 3:1
to about 5:1, about 5:1 to about 7:1, about 7:1 to about 9:1, about
9:1 to about 11:1, about 2:1 to about 4:1, about 4:1 to about 6:1,
about 6:1 to about 8:1, about 8:1 to about 12:1, about 12:1 to
about 14:1, about 14:1 to about 16:1, about 16:1 to about 18:1,
about 3:1 to about 7:1, about 7:1 to about 11:1, about 11:1 to
about 15:1, about 15:1 to about 19:1, about 4:1 to about 8:1, about
8:1 to about 12:1, about 12:1 to about 16:1, or about 16:1 to about
20:1.
In some embodiments, to determine the weight ratio of the total
albumin to the total rapamycin in the composition (such as a
pharmaceutical composition), the amount of the albumin on the
nanoparticle, the amount of the albumin not on the nanoparticle
(e.g., in the non-nanoparticle portion, or free in solution in the
composition (such as a pharmaceutical composition)), the amount of
the rapamycin in the nanoparticle, and the amount of the rapamycin
not associated with the nanoparticle (e.g., in the non-nanoparticle
portion, or free in solution in the composition (such as a
pharmaceutical composition)) are needed. As discussed previously,
the amount of the albumin on the nanoparticles can be determined
by, for example, chromatography, such as size-exclusion
chromatography, or spectrophotometric measurements following
isolation of the nanoparticles in the pharmaceutical composition.
In some embodiments, for example, following ultracentrifugation to
pellet the nanoparticles, the amount of albumin in the resulting
supernatant can be determined by similar methods discussed above
for determining the amount of the albumin on the nanoparticles. As
discussed previously, the amount of the rapamycin in the
nanoparticles can be determined by, for example, chromatography,
such as RP-HPLC, spectrophotometric measurements, or mass
spectrometric measurements. In some embodiments, for example,
following ultracentrifugation to pellet the nanoparticles, the
amount of rapamycin in the resulting supernatant can be determined
by similar methods discussed above for determining the amount of
rapamycin not on the nanoparticles.
In some embodiments, to determine the weight ratio of the total
albumin to the total rapamycin in the composition (such as a
pharmaceutical composition), the amount of the total albumin in the
composition (such as a pharmaceutical composition) and the amount
of the total rapamycin in the composition (such as a pharmaceutical
composition) are needed. The amount of the total albumin, for
example, can be measured using the methods discussed above using a
sample that contains all albumins from the composition (such as a
pharmaceutical composition). The amount of the total rapamycin, for
example, can be measured using the methods discussed above using a
sample that contains all rapamycin from the composition (such as a
pharmaceutical composition).
The weight ratio of the total albumin to the total poorly water
soluble drug (such as rapamycin) in the composition (such as a
pharmaceutical composition) can be determined, for example, by
calculating the ratio of the amount of the total albumin in the
composition (such as a pharmaceutical composition) and the amount
of the total rapamycin in the composition (such as a pharmaceutical
composition).
Poorly Water Soluble Drugs
Poorly water soluble drugs described herein can be, for example,
drugs with solubility in water less than about 10 mg/ml at about
20-25.degree. C., including for example drugs with solubility less
than about any of 5 mg/ml, 2 mg/ml, 1 mg/ml, 0.5 mg/ml, 0.2 mg/ml,
0.1 mg/ml, 0.05 mg/ml, 0.02 mg/ml, or 0.01 mg/ml. In some
embodiments, the poorly water soluble drug is an antineoplastic
agent. In some embodiments, the poorly water soluble drug is a
chemotherapeutic agent. Suitable poorly water soluble drugs
include, but are not limited to, taxanes (such as paclitaxel,
docetaxel, ortataxel and other taxanes), 17-allyl amino
geldanamycin (17-AAG), or thiocolchicine dimer (such as
IDN5404).
In some embodiments, the poorly water soluble drug is a limus drug,
which includes rapamycin (sirolimus) and its analogues. Examples of
limus drugs include, but are not limited to, temsirolimus
(CCI-779), everolimus (RAD001), ridaforolimus (AP-23573),
deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and
tacrolimus (FK-506). In some embodiments, the limus drug is
selected from the group consisting of temsirolimus (CCI-779),
everolimus (RAD001), ridaforolimus (AP-23573), deforolimus
(MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus
(FK-506). In some embodiments, the poorly water soluble drug is
rapamycin (sirolimus).
Other Components in the Albumin-Based Nanoparticle Composition
The compositions (such as pharmaceutical compositions) described
herein may also include an antimicrobial agent (e.g., an agent in
addition to the rapamycin) in an amount sufficient to significantly
inhibit (e.g., delay, reduce, slow, and/or prevent) microbial
growth in the composition (such as a pharmaceutical composition)
for use in the methods of treatment, methods of administration, and
dosage regimens described herein. Exemplary microbial agents and
variations for the use of microbial agents are disclosed in US
2007/0117744 A1 (such as those described in paragraphs [0036] to
[0058] therein), the content of which is hereby incorporated by
reference in its entirety. In some embodiments, the antimicrobial
agent is a chelating agent, such as EDTA, edetate, citrate,
pentetate, tromethamine, sorbate, ascorbate, derivatives thereof,
or mixtures thereof. In some embodiments, the antimicrobial agent
is a polydentate chelating agent. In some embodiments, the
antimicrobial agent is a non-chelating agent, such as any of
sulfites, benzoic acid, benzyl alcohol, chlorobutanol, and paraben.
In some embodiments, an antimicrobial other than the taxane
discussed above is not contained or used in the methods of
treatment, methods of administration, and dosage regimens described
herein.
In some embodiments, the compositions (such as pharmaceutical
compositions) described herein include a sugar. Exemplary sugars
and variations for the use of sugars are disclosed in US
2007/0117744 A1 (such as those described in paragraphs [0084] to
[0090] therein), the content of which is hereby incorporated by
reference in its entirety. In some embodiments, the sugar serves as
a reconstitution enhancer which causes a lyophilized composition to
dissolve or suspend in water and/or aqueous solution more quickly
than the lyophilized composition would dissolve without the sugar.
In some embodiments, the composition (such as a pharmaceutical
composition) is a liquid (e.g., aqueous) composition obtained by
reconstituting or resuspending a dry composition. In some
embodiments, the concentration of sugar in the composition (such as
a pharmaceutical composition) is greater than about 50 mg/ml. In
some embodiments, the sugar is in an amount that is effective to
increase the stability of the rapamycin in the composition (such as
a pharmaceutical composition) as compared to a composition (such as
a pharmaceutical composition) without the sugar. In some
embodiments, the sugar is in an amount that is effective to improve
filterability of the composition (such as a pharmaceutical
composition) as compared to a composition (such as a pharmaceutical
composition) without the sugar.
The sugar-containing compositions (such as pharmaceutical
compositions) described herein may further comprise one or more
antimicrobial agents, such as the antimicrobial agents described
herein or in US 2007/0117744 A1. In addition to one or more sugars,
other reconstitution enhancers (such as those described in US
2005/0152979 A1, which is hereby incorporated by reference in its
entirety) can also be added to the compositions (such as
pharmaceutical compositions).
The compositions (such as pharmaceutical compositions) described
herein may be used in pharmaceutical compositions or formulations,
by combining the pharmaceutical composition(s) described with a
pharmaceutically acceptable carrier, excipients, stabilizing agents
and/or other agents, which are known in the art, for use in the
methods of treatment, methods of administration, and dosage
regimens described herein.
To increase stability by increasing the negative zeta-potential of
nanoparticles, certain negatively charged components may be added.
Such negatively charged components include, but are not limited to
bile salts, bile acids, glycocholic acid, cholic acid,
chenodeoxycholic acid, taurocholic acid, glycochenodeoxycholic
acid, taurochenodeoxycholic acid, litocholic acid, ursodeoxycholic
acid, dehydrocholic acid, and others; phospholipids including
lecithin (egg yolk) based phospholipids which include the following
phosphatidylcholines: palmitoyloleoylphosphatidylcholine,
palmitoyllinoleoylphosphatidylcholine,
stearoyllinoleoylphosphatidylcholine,
stearoyloleoylphosphatidylcholine,
stearoylarachidoylphosphatidylcholine, and
dipalmitoylphosphatidylcholine. Other phospholipids including
L-.alpha.-dimyristoylphosphatidylcholine (DMPC),
dioleoylphosphatidylcholine (DOPC), distearoylphosphatidylcholine
(DSPC), hydrogenated soy phosphatidylcholine (HSPC), and other
related compounds. Negatively charged surfactants or emulsifiers
are also suitable as additives, e.g., sodium cholesteryl sulfate
and the like.
Suitable pharmaceutical carriers include sterile water; saline,
dextrose; dextrose in water or saline; condensation products of
castor oil and ethylene oxide combining about 30 to about 35 moles
of ethylene oxide per mole of castor oil; liquid acid; lower
alkanols; oils such as corn oil; peanut oil, sesame oil and the
like, with emulsifiers such as mono- or di-glyceride of a fatty
acid, or a phosphatide, e.g., lecithin, and the like; glycols;
polyalkylene glycols; aqueous media in the presence of a suspending
agent, for example, sodium carboxymethylcellulose; sodium alginate;
poly(vinylpyrrolidone); and the like, alone, or with suitable
dispensing agents such as lecithin; polyoxyethylene stearate; and
the like. The carrier may also contain adjuvants such as preserving
stabilizing, wetting, emulsifying agents and the like together with
the penetration enhancer. The final form may be sterile and may
also be able to pass readily through an injection device such as a
hollow needle. The proper viscosity may be achieved and maintained
by the proper choice of solvents or excipients. Moreover, the use
of molecular or particulate coatings such as lecithin, the proper
selection of particle size in dispersions, or the use of materials
with surfactant properties may be utilized.
The pharmaceutical compositions described herein may include other
agents, excipients, or stabilizers to improve properties of the
composition (such as a pharmaceutical composition). Examples of
suitable excipients and diluents include, but are not limited to,
lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum
acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium
silicate, microcrystalline cellulose, polyvinylpyrrolidone,
cellulose, water, saline solution, syrup, methylcellulose, methyl-
and propylhydroxybenzoates, talc, magnesium stearate and mineral
oil. The formulations can additionally include lubricating agents,
wetting agents, emulsifying and suspending agents, preserving
agents, sweetening agents or flavoring agents. Examples of
emulsifying agents include tocopherol esters such as tocopheryl
polyethylene glycol succinate and the like, copolymer emulsifier
sold under the trademark PLURONIC.RTM., emulsifiers based on
polyoxy ethylene compounds, nonionic surfactant sold under the
trademark SPAN.RTM. 80 and related compounds and other emulsifiers
known in the art and approved for use in animals or human dosage
forms. The compositions (such as pharmaceutical compositions) can
be formulated so as to provide rapid, sustained or delayed release
of the active ingredient after administration to the patient by
employing procedures well known in the art.
In some embodiments, the composition (such as a pharmaceutical
composition) is formulated to have a pH in the range of about 4.5
to about 9.0, including for example pH ranges of any one of about
5.0 to about 8.0, about 6.5 to about 7.5, or about 6.5 to about
7.0. In some embodiments, the pH of the composition (such as a
pharmaceutical composition) is formulated to no less than about 6,
including for example no less than about any one of 6.5, 7, or 8
(e.g., about 8). The composition (such as a pharmaceutical
composition) can also be made to be isotonic with blood by the
addition of a suitable tonicity modifier, such as glycerol.
In some embodiments, the composition (such as a pharmaceutical
composition) is suitable for administration to a human. In some
embodiments, the composition (such as a pharmaceutical composition)
is suitable for administration to a human by parenteral
administration. Formulations suitable for parenteral administration
include aqueous and non-aqueous, isotonic sterile injection
solutions, which can contain anti-oxidants, buffers, bacteriostats,
and solutes that render the formulation compatible with the blood
of the intended recipient, and aqueous and non-aqueous sterile
suspensions that can include suspending agents, solubilizers,
thickening agents, stabilizing agents, and preservatives. The
formulations can be presented in unit-dose or multi-dose sealed
containers, such as ampules and vials, and can be stored in a
freeze-dried (lyophilized) condition requiring only the addition of
the sterile liquid excipient methods of treatment, methods of
administration, and dosage regimens described herein (e.g., water)
for injection, immediately prior to use. Extemporaneous injection
solutions and suspensions can be prepared from sterile powders,
granules, and tablets of the kind previously described. Injectable
formulations are preferred. In some embodiments, the composition
(such as a pharmaceutical composition) is contained in a single-use
vial, such as a single-use sealed vial. In some embodiments, each
single-use vial contains about 100 mg rapamycin. In some
embodiments, the single-use vial contains about 900 mg albumin. In
some embodiments, the composition (such as a pharmaceutical
composition) is contained in a multi-use vial. In some embodiments,
the composition (such as a pharmaceutical composition) is contained
in bulk in a container.
Also provided are unit dosage forms comprising the compositions
(such as pharmaceutical compositions) and formulations described
herein. These unit dosage forms can be stored in a suitable
packaging in single or multiple unit dosages and may also be
further sterilized and sealed. In some embodiments, the composition
(such as a pharmaceutical composition) (such as a pharmaceutical
composition) also includes one or more other compounds (or
pharmaceutically acceptable salts thereof) that are useful for
treating cancer. In various variations, the amount of rapamycin in
the composition (such as a pharmaceutical composition) is included
in any one of the following ranges: about 5 mg to about 50 mg,
about 20 mg to about 50 mg, about 50 mg to about 100 mg, about 100
mg to about 125 mg, about 125 mg to about 150 mg, about 150 mg to
about 175 mg, about 175 mg to about 200 mg, about 200 mg to about
225 mg, about 225 mg to about 250 mg, about 250 mg to about 300 mg,
about 300 mg to about 350 mg, about 350 mg to about 400 mg, about
400 mg to about 450 mg, or about 450 mg to about 500 mg. In some
embodiments, the amount of rapamycin in the composition (such as a
pharmaceutical composition) (e.g., a dosage or unit dosage form) is
in the range of about 5 mg to about 500 mg, such as about 30 mg to
about 300 mg or about 50 mg to about 200 mg. In some embodiments,
the carrier is suitable for parental administration (e.g.,
intravenous administration). In some embodiments, the rapamycin is
the only pharmaceutically active agent for the treatment of cancer
that is contained in the composition (such as a pharmaceutical
composition).
In some embodiments, there is provided a dosage form (e.g., a unit
dosage form) for the treatment of cancer comprising any one of the
compositions (such as pharmaceutical compositions) described
herein. In some embodiments, there are provided articles of
manufacture comprising the compositions (such as pharmaceutical
compositions), formulations, and unit dosages described herein in
suitable packaging for use in the methods of treatment, methods of
administration, and dosage regimens described herein. Suitable
packaging for compositions (such as pharmaceutical compositions)
described herein are known in the art, and include, for example,
vials (such as sealed vials), vessels (such as sealed vessels),
ampules, bottles, jars, flexible packaging (e.g., sealed bags sold
under the trademark MYLAR.RTM. or plastic bags), and the like.
These articles of manufacture may further be sterilized and/or
sealed.
Methods of Validating and/or Releasing a Commercial Batch of
Albumin-Based Nanoparticle Compositions
The methods described herein may be carried out when validating
and/or releasing a commercial batch of an albumin-based
nanoparticle composition. "Commercial batch" used herein refers to
a batch size that is at least about 20 grams (by amount of poorly
water soluble drug (such as rapamycin)). In some embodiments, the
batch size is at least about 30 g, 40 g, 50 g, 60 g, 70 g, 80 g, 90
g, 100 g, 150 g, 200 g, 250 g, 300 g, 350 g, 400 g, 450 g, 500 g,
550 g, 600 g, 650 g, 700 g, 750 g, 800 g, 850 g, 900 g, 1000 g,
1500 g, 2000 g, 2500 g, 3000 g, 3500 g, 4000 g, 4500 g, 5000 g, or
10,000 grams (by amount of poorly water soluble drug (such as
rapamycin)). In some embodiments, the commercial batch comprises a
plurality of vials comprising any of the compositions (such as
pharmaceutical compositions) described herein. In some embodiments,
the commercial batch comprises at least about any of 100 vials, 150
vials, 200 vials, 100 vials, 150 vials, 200 vials, 250 vials, 300
vials, 350 vials, 400 vials, 450 vials, 500 vials, 550 vials, 600
vials, 650 vials, 700 vials, 750 vials, 800 vials, 850 vials, 900
vials, 1000 vials, 1500 vials, 2000 vials, 2500 vials, 3000 vials,
3500 vials, 4000 vials, 4500 vials, 5000 vials, 10000 vials, 12000
vials, 14000 vials, 16000 vials, 18000 vials, 20000 vials, 22000
vials, 24000 vials, 26000 vials, 28000 vials, 30000 vials, 32000
vials, 34000 vials, 36000 vials, 38000 vials, 40000 vials, 42000
vials, 44000 vials, 46000 vials, 48000 vials, or 50000 vials. For
example, each vial contains about any of 100 mg, 200 mg, 300 mg,
400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or 1000 mg of the
composition (such as a pharmaceutical composition). In some
embodiments, the pharmaceutical composition in the commercial batch
is a liquid suspension. In some embodiments, the pharmaceutical
composition in the commercial batch is a lyophilized powder.
The present application thus in some embodiments provides a method
of validating and/or releasing a commercial batch of a
pharmaceutical composition for medical use in a human individual,
wherein the pharmaceutical composition comprise nanoparticles
comprising rapamycin coated with albumin and a non-nanoparticle
portion comprising albumin and rapamycin, the method comprises a)
obtaining a sample from the commercial batch; and 2) assessing
suitability of the pharmaceutical composition for medical use
according to any one of the methods described herein. In some
embodiments, at least about 2 samples, 3 samples, 4 samples, 5
samples, 6 samples, 10 samples, 20 samples, 30 samples, 40 samples,
50 samples, 60 samples, 70 samples, 80 samples, 90 samples, 100
samples, or more samples are obtained from the commercial batch and
subject to assessment. In some embodiments, the amount of the
sample (i.e., the amount of the pharmaceutical composition taken
from the commercial batch) is about any of 10-20 .mu.g, 20-50
.mu.g, 50-100 .mu.g, 100-200 .mu.g, 200-500 .mu.g, 500-1000 .mu.g,
1000 .mu.g, 2000 .mu.g, 3000 .mu.g, 4000 .mu.g, 5000 .mu.g, or
greater than 5000 .mu.g. In some embodiments, the sample is the
pharmaceutical composition in a vial. In some embodiments, the
sample is obtained from the commercial batch prior to
lyophilization of the composition (such as a pharmaceutical
composition). In some embodiments, the sample is obtained from the
commercial batch after lyophilization of the composition (such as a
pharmaceutical composition). In some embodiments, the sample is
obtained from the commercial batch after reconstitution.
Thus, for example, the present application in some embodiments
provides a method of validating and/or releasing a commercial batch
of a pharmaceutical composition for medical use in a human
individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, the
method comprising: a) obtaining a sample from the commercial batch;
and b) determining the percentage of albumin polymers among the
albumin on the nanoparticles in the sample, wherein a percentage of
albumin polymer among the albumin on the nanoparticles being about
15% to about 40% (such as about 15% to about 20%, about 20% to
about 24.5%, about 24.5% to about 30%, about 30% to about 35%, or
about 35% to about 40%) is indicative of suitability of the
commercial batch for medical use and/or release.
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin, the method comprising: a) obtaining a sample from the
commercial batch; and b) determining the percentage of albumin
monomers among the albumin on the nanoparticles in the sample,
wherein a percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%) is indicative of suitability of the
commercial batch for medical use and/or release.
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin, the method comprising: a) obtaining a sample from the
commercial batch; and b) determining the weight percentage of the
albumin in the nanoparticles, wherein a weight percentage of the
albumin in the nanoparticles being about 15% to about 30% (such as
about 20% to about 25%, about 15% to about 24%, or about 15% to
about 20%) is indicative of suitability of the commercial batch for
medical use and/or release.
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin, the method comprising: a) obtaining a sample from the
commercial batch; and b) determining the weight ratio of albumin to
rapamycin in the nanoparticles, wherein an albumin to rapamycin
ratio of about 1:2 to about 1:6 in the nanoparticles is indicative
of suitability of the commercial batch for medical use and/or
release.
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin, the method comprising: a) obtaining a sample from the
commercial batch; and b) determining the morphology of the
nanoparticles under cryo-TEM, wherein an irregular shape of the
nanoparticles is indicative of suitability of the commercial batch
for medical use and/or release.
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin, the method comprising: a) obtaining a sample from the
commercial batch; and b) determining the thickness of the albumin
coating of the nanoparticles under cryo-TEM, wherein a thickness of
about 5-7 nm (such as about 6 nm) is indicative of suitability of
the commercial batch for medical use and/or release.
In some embodiments, there is provided a method of validating
and/or release a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin, the method comprising: a) obtaining a sample from the
commercial batch; and b) determining the distribution of rapamycin
in a tumor tissue upon injection of the pharmaceutical composition
directly into the tumor tissue; wherein an enhanced rapamycin tumor
distribution is indicative of suitability of the commercial batch
for medical use and/or release.
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin, the method comprising: a) obtaining a sample from the
commercial batch; and b) determining the solubility, rapamycin
crystallinity, and a recovery following a 0.2 micron filtration of
the pharmaceutical composition, wherein a solubility of about 50
.mu.g/ml to about 100 .mu.g/ml in a 5% human albumin solution, a
non-crystalline state of the rapamycin, and a recovery date of at
least about 80% is indicative of suitability of the commercial
batch for medical use and/or release.
In some embodiments, the method comprises various combinations of
the determination steps described above. In some embodiments, the
method comprises at least 2, 3, 4, 5, 6, 7, or 8 determination
steps described above. In some embodiments, the method further
comprises determining at least one (such as at least any of 2, 3,
4, 5, 6, 7, 8, 9, or 10) of the following characteristics of the
sample: 1) binding affinity of albumin to rapamycin in the
composition (such as a pharmaceutical composition) (for example by
equilibrium dialysis, FTIR, NMR, or a combination thereof); 2)
surface-to-volume ratio; 3) percentage of albumin dimers and/or
oligomers among the albumin on the nanoparticles; 4) distribution
of the total rapamycin and/or the total albumin between the
nanoparticles and the non-nanoparticle portion; 5) oligomeric
status of the total albumin in the composition (such as a
pharmaceutical composition); 6) particle size of the nanoparticles,
including average particle size, polydispersity, and/or size
distribution; 7) surface potential; 8) in vitro release kinetics;
9) physical stability; and 10) rapamycin tumor distribution in
vivo. In some embodiments, at least one (such as at least any of 2,
3, 4, 5, 6, 7, 8, or 9) of the determination steps are carried out
upon reconstitution of the sample. In some embodiments, at least
one (such as at least any of 2, 3, 4, 5, 6, 7, 8, or 9) of the
determination steps are carried out upon storage of the sample. In
some embodiments, at least one of the determination steps are
carried out after storage of the sample for at least about any of 6
hours, 12 hours, 24 hours, 36 hours, 48 hours, or 72 hours. In some
embodiments, at least one of the determination steps are carried
out after storage of the sample at room temperature, under
refrigerated condition, or at about 40.degree. C.
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin, the method comprising: determining the solubility,
rapamycin crystallinity, and rapamycin recovery following a 0.2
micron filtration of the pharmaceutical composition, determining
the percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition,
determining the percentage of the rapamycin in the nanoparticles
among the total rapamycin in the pharmaceutical composition (for
example by reversed-phase HPLC), determining the percentage of the
albumin that is in the non-nanoparticle portion among the total
albumin in the pharmaceutical composition (for example by
size-exclusion chromatography), determining the particle size of
the nanoparticles (for example by dynamic light scattering) and/or
size distribution of the nanoparticles, and determining the
stability of the pharmaceutical composition (including determining
stability after storage). In some embodiments, the solubility,
rapamycin crystallinity, and/or rapamycin recovery are determined
after storage (for example after storage for at least about any of
6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 hours, or 72
hours, such as at room temperature, under refrigerated condition,
or at about 40.degree. C.). In some embodiments, the rapamycin
crystallinity is determined by X-ray diffraction and/or polarized
light microscopy. In some embodiments, the method further comprises
determining binding affinity of albumin to rapamycin in the
composition (such as a pharmaceutical composition) (for example by
equilibrium dialysis, FTIR, NMR, or a combination thereof). In some
embodiments, the method further comprises determining tumor
distribution of rapamycin upon administration in vivo (for example
by determining tumor distribution of rapamycin upon injection of
the pharmaceutical composition directly into the tumor tissue).
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the commercial batch
of a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein the nanoparticles having less than
about 52% of the albumin on the nanoparticles in the form of
monomers is indicative of suitability of the commercial batch for
medical use and/or release. In some embodiments, there is provided
a method of validating and/or releasing a commercial batch of a
pharmaceutical composition for medical use in a human individual,
wherein the commercial batch of a pharmaceutical composition
comprising a pharmaceutical composition comprising: a)
nanoparticles comprising rapamycin coated with albumin, and b) a
non-nanoparticle portion comprising albumin and rapamycin, wherein
the nanoparticles having more than about 35% of albumin on the
nanoparticles in the forms of polymers and oligomers is indicative
of suitability of the commercial batch for medical use and/or
release. In some embodiments, there is provided a method of
validating and/or releasing a commercial batch of a pharmaceutical
composition for medical use in a human individual, wherein the
commercial batch of a pharmaceutical composition comprising a
pharmaceutical composition comprising: a) nanoparticles comprising
rapamycin coated with albumin, and b) a non-nanoparticle portion
comprising albumin and rapamycin, wherein the nanoparticles having
less than about 54% of the albumin on the nanoparticles in the form
of monomers, and more than about 11% of the albumin in the
nanoparticles in the form of polymers is indicative of suitability
of the commercial batch for medical use and/or release. In some
embodiments, there is provided a method of validating and/or
releasing a commercial batch of a pharmaceutical composition for
medical use in a human individual, wherein the commercial batch of
a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein the nanoparticles having less than
about 55% of the albumin on the nanoparticles in the form of
monomers, and more than about 18% of the albumin in the
nanoparticles in the form of polymers is indicative of suitability
of the commercial batch for medical use and/or release. In some
embodiments, there is provided a method of validating and/or
releasing a commercial batch of a pharmaceutical composition for
medical use in a human individual, wherein the commercial batch of
a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein the nanoparticles having a ratio of
albumin on the nanoparticles in the form of polymers and oligomers
divided by the albumin on the nanoparticles in the form of monomers
of more than about 62% is indicative of suitability of the
commercial batch for medical use and/or release. In some
embodiments, there is provided a method of validating and/or
releasing a commercial batch of a pharmaceutical composition for
medical use in a human individual, wherein the commercial batch of
a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein the nanoparticles having a weight
percentage of the albumin in the nanoparticles of about 15% to
about 30% (such as about 20% to about 25%, about 15% to about 24%,
or about 15% to about 20%) is indicative of suitability of the
commercial batch for medical use and/or release. In some
embodiments, there is provided a method of validating and/or
releasing a commercial batch of a pharmaceutical composition for
medical use in a human individual, wherein the commercial batch of
a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein the nanoparticles having a weight
ratio of the albumin to the rapamycin in the nanoparticles of about
1:2 to about 1:6 is indicative of suitability of the commercial
batch for medical use and/or release. In some embodiments, there is
provided a method of validating and/or releasing a commercial batch
of a pharmaceutical composition for medical use in a human
individual, wherein the commercial batch of a pharmaceutical
composition comprising a pharmaceutical composition comprising: a)
nanoparticles comprising rapamycin coated with albumin, and b) a
non-nanoparticle portion comprising albumin and rapamycin, wherein
the nanoparticles in the pharmaceutical composition having an
irregular shape is indicative of suitability of the commercial
batch for medical use and/or release. In some embodiments, there is
provided a method of validating and/or releasing a commercial batch
of a pharmaceutical composition for medical use in a human
individual, wherein the commercial batch of a pharmaceutical
composition comprising a pharmaceutical composition comprising: a)
nanoparticles comprising rapamycin coated with albumin, and b) a
non-nanoparticle portion comprising albumin and rapamycin, wherein
a thickness of the albumin coating on the nanoparticles of about
5-7 nm as measured by cryo-TEM is indicative of suitability of the
commercial batch for medical use and/or release. In some
embodiments, there is provided a method of validating and/or
releasing a commercial batch of a pharmaceutical composition for
medical use in a human individual, wherein the commercial batch of
a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein a weight ratio of the total albumin
to the total rapamycin in the pharmaceutical composition of about
3:1 to about 7.9:1 or about 10:1 to about 17:1 is indicative of
suitability of the commercial batch for medical use and/or release.
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the commercial batch
of a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein a solubility of about 50 .mu.g/ml to
about 100 .mu.g/ml (including for example any of about 50 .mu.g/ml
to about 60 .mu.g/ml, about 60 .mu.g/ml to about 70 .mu.g/ml, about
70 .mu.g/ml to about 75 .mu.g/ml, about 75 .mu.g/ml to about 80
.mu.g/ml, about 80 .mu.g/ml to about 90 .mu.g/ml, or about 90
.mu.g/ml to about 100 .mu.g/ml) when diluted in a 5% human albumin
solution is indicative of suitability of the commercial batch for
medical use and/or release. In some embodiments, there is provided
a method of validating and/or releasing a commercial batch of a
pharmaceutical composition for medical use in a human individual,
wherein the commercial batch of a pharmaceutical composition
comprising a pharmaceutical composition comprising: a)
nanoparticles comprising rapamycin coated with albumin, and b) a
non-nanoparticle portion comprising albumin and rapamycin, wherein
a rapamycin recovery of at least about 80% (including for example
at least about any of 85%, 90%, 95%, 98%, or 99%) following a 0.2
micron filtration is indicative of suitability of the commercial
batch for medical use and/or release. In some embodiments, there is
provided a method of validating and/or releasing a commercial batch
of a pharmaceutical composition for medical use in a human
individual, wherein the commercial batch of a pharmaceutical
composition comprising a pharmaceutical composition comprising: a)
nanoparticles comprising rapamycin coated with albumin, and b) a
non-nanoparticle portion comprising albumin and rapamycin, wherein
an average particle size of the nanoparticles in the pharmaceutical
composition of less than about 200 nm (including for example about
100 nm to about 160 nm) is indicative of suitability of the
commercial batch for medical use and/or release. In some
embodiments, there is provided a method of validating and/or
releasing a commercial batch of a pharmaceutical composition for
medical use in a human individual, wherein the commercial batch of
a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein the nanoparticles in the
pharmaceutical composition having a zeta-potential of about -20 mV
to about -35 mV is indicative of suitability of the commercial
batch for medical use and/or release. In some embodiments, there is
provided a method of validating and/or releasing a commercial batch
of a pharmaceutical composition for medical use in a human
individual, wherein the commercial batch of a pharmaceutical
composition comprising a pharmaceutical composition comprising: a)
nanoparticles comprising rapamycin coated with albumin, and b) a
non-nanoparticle portion comprising albumin and rapamycin, wherein
the nanoparticles in the pharmaceutical composition having a
polydispersity index of less than about 0.3 (including for example
less than about 0.06 or about any of 0.05-0.09, 0.09-0.13,
0.13-0.15, 0.15-0.2, or 0.2-0.3) is indicative of suitability of
the commercial batch for medical use and/or release. In some
embodiments, there is provided a method of validating and/or
releasing a commercial batch of a pharmaceutical composition for
medical use in a human individual, wherein the commercial batch of
a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein the nanoparticles in the
pharmaceutical composition having a span of size distribution
((Dv90-Dv10)/Dv50) of about 0.8 to about 1.5 (including for example
about any of 0.8-0.9, 0.9-1, 1-1.1, 1.1-1.2, 1.2-1.3, 1.3-1.4, or
1.4-1.5) is indicative of suitability of the commercial batch for
medical use and/or release. In some embodiments, there is provided
a method of validating and/or releasing a commercial batch of a
pharmaceutical composition for medical use in a human individual,
wherein the commercial batch of a pharmaceutical composition
comprising a pharmaceutical composition comprising: a)
nanoparticles comprising rapamycin coated with albumin, and b) a
non-nanoparticle portion comprising albumin and rapamycin, wherein
the rapamycin in the pharmaceutical composition being
non-crystalline (including for example non-crystalline after
storage for about 24 hours at about 4.degree. C.) is indicative of
suitability of the commercial batch for medical use and/or release.
In some embodiments, there is provided a method of validating
and/or releasing a commercial batch of a pharmaceutical composition
for medical use in a human individual, wherein the commercial batch
of a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein the pharmaceutical composition upon
tumor injection allowing rapamycin to spread radially for a
distance of greater than (for example more than about any of
1.1.times., 1.2.times., 1.3.times., 1.4.times., 1.5.times.,
1.6.times., 1.7.times., 1.8.times., 2.times., 3.times., 4.times.,
5.times., 6.times., 7.times., 8.times., or more of) that of a
solvent-based rapamycin formulation (such as the solvent-based
paclitaxel formulation sold under the trademark TAXOL.RTM.) under
the same assay conditions is indicative of suitability of the
commercial batch for medical use and/or release. In some
embodiments, there is provided a method of validating and/or
releasing a commercial batch of a pharmaceutical composition for
medical use in a human individual, wherein the commercial batch of
a pharmaceutical composition comprising a pharmaceutical
composition comprising: a) nanoparticles comprising rapamycin
coated with albumin, and b) a non-nanoparticle portion comprising
albumin and rapamycin, wherein the pharmaceutical composition upon
tumor injection allowing rapamycin to spread radically for more
than about 700 .mu.m (such as more than about any of 700 .mu.m, 800
.mu.m, 900 .mu.m, 1000 .mu.m, 1100 .mu.m or 1200 .mu.m) within
about 24 hours after the pharmaceutical composition is injected
into a tumor tissue (for example injected at the rapamycin amount
of about 12 .mu.g (such as at about 4 mg/ml) into a pancreatic MIA
PaCa-2 xenograft tumor) is indicative of suitability of the
commercial batch for medical use and/or release.
Also provided are commercial batches released by following any one
of the methods described herein.
Further provided are kits for use in any one of the methods of
assessment and commercial batch release described herein.
Kits Comprising the Albumin-Based Nanoparticle Compositions
Once determined to be suitable for medical use, the pharmaceutical
compositions can be included in kits comprising the compositions
(such as pharmaceutical compositions), formulations, unit dosages,
and articles of manufacture for use in the methods of treatment,
methods of administration, and dosage regimens described herein.
Kits described herein include one or more containers comprising the
rapamycin pharmaceutical compositions (formulations or unit dosage
forms and/or articles of manufacture), and in some embodiments,
further comprise instructions for accessing and/or using in
accordance with any of the methods of treatment described herein.
In various embodiments, the amount of rapamycin in the kit is
included in any one of the following ranges: about 5 mg to about 20
mg, about 20 mg to about 50 mg, about 50 mg to about 100 mg, about
100 mg to about 125 mg, about 125 mg to about 150 mg, about 150 mg
to about 175 mg, about 175 mg to about 200 mg, about 200 mg to
about 225 mg, about 225 mg to about 250 mg, about 250 mg to about
300 mg, about 300 mg to about 350 mg, about 350 mg to about 400 mg,
about 400 mg to about 450 mg, or about 450 mg to about 500 mg. In
some embodiments, the amount of rapamycin in the kit is in the
range of about 5 mg to about 500 mg, such as about 30 mg to about
300 mg or about 50 mg to about 200 mg. In some embodiments, the kit
includes one or more other compounds (e.g., one or more compounds
other than rapamycin that are useful for cancer).
Instructions supplied in the kits described herein are typically
written instructions on a label or package (e.g., a paper sheet
included in the kit), but machine-readable instructions (e.g.,
instructions carried on a magnetic or optical storage disk) are
also acceptable. The instructions relating to the use of the
pharmaceutical compositions generally include information as to
dosage, dosing schedule, and route of administration for the
intended treatment. The kit may further comprise a description of
selecting an individual suitable or treatment.
The present application also provides kits comprising compositions
(such as pharmaceutical compositions) (or unit dosages forms and/or
articles of manufacture) described herein and may further comprise
instruction(s) on methods of assessing and/or using the composition
(such as a pharmaceutical composition), such as uses further
described herein. In some embodiments, the kit described herein
comprises the packaging described above. In other variations, the
kit described herein comprises the packaging described above and a
second packaging comprising a buffer. It may further include other
materials desirable from a commercial and user standpoint,
including other buffers, diluents, filters, needles, syringes, and
packages with instructions for performing any methods described
herein.
For combination therapies described herein, the kit may contain
instructions for administering the first and second therapies
simultaneously and/or sequentially for the effective treatment of
cancer. The first and second therapies can be present in separate
containers or in a single container. It is understood that the kit
may comprise one distinct composition or two or more compositions
(such as pharmaceutical compositions) wherein one composition
comprises a first therapy and one composition comprises a second
therapy.
Kits may also be provided that contain sufficient dosages of the
rapamycin as disclosed herein to provide effective treatment for an
individual for an extended period, such as any one of a week, 2
weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5
months, 6 months, 7 months, 8 months, 9 months or more. Kits may
also include multiple unit doses of the rapamycin, compositions
(such as pharmaceutical compositions), and formulations described
herein and instructions for use and packaged in quantities
sufficient for storage and use in pharmacies, for example, hospital
pharmacies and compounding pharmacies. In some embodiments, the kit
comprises a dry (e.g., lyophilized) composition that can be
reconstituted, resuspended, or rehydrated to form generally a
stable aqueous suspension of nanoparticles comprising rapamycin and
albumin.
The kits described herein are in suitable packaging. Suitable
packaging include, but is not limited to, vials, bottles, jars,
flexible packaging (e.g., sealed bags sold under the trademark
MYLAR.RTM. or plastic bags), and the like. Kits may optionally
provide additional components such as buffers and interpretative
information.
Methods of Making the Pharmaceutical Compositions
The present application also provides methods of making the
pharmaceutical compositions described herein. Nanoparticles
containing poorly water soluble pharmaceutical agents and carrier
proteins (e.g., albumin) can be prepared under conditions of high
shear forces (e.g., sonication, high pressure homogenization, or
the like). These methods are disclosed in, for example, U.S. Pat.
Nos. 5,916,596 A; 6,096,331 A; 6,749,868 B1; 6,537,579 B1; and WO
1998/014174 A1; WO 1999/000113 A1; WO 2007/027941 A2; and WO
2007/027819 A2. The contents of these publications, particularly
with respect to the method of making compositions (such as
pharmaceutical compositions) containing carrier proteins, are
hereby incorporated by reference in their entireties.
Generally, to make the rapamycin pharmaceutical compositions
described herein, rapamycin is dissolved in an organic solvent.
Suitable organic solvents include, for example, ketones, esters,
ethers, chlorinated solvents, and other solvents known in the art.
For example, the organic solvent can be methylene chloride/ethanol,
chloroform/ethanol, or chloroform/t-butanol (for example with a
ratio of about any one of 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2,
1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, or 9:1 or with a ratio of
about any one of 3:7, 5:7, 4:6, 5:5, 6:5, 8:5, 9:5, 9.5:5, 5:3,
7:3, 6:4, or 9.5:0.5). Albumin (such as recombinant albumin, for
example recombinant albumin sold under the trademark NOVOZYMES.RTM.
or recombinant albumin sold under the trademark INVITRIA.TM.
disclosed herein) is dissolved in water and combined with the
rapamycin solution. The mixture is subjected to high pressure
homogenization (e.g., using a high pressure homogenizer sold by
Avestin, APV Gaulin, or Stansted, or a high pressure homogenizer
sold under the trademark MICROFLUIDIZER.TM. such as the high
pressure homogenizer sold under the trademark MICROFLUIDIZER.TM.
Processor M-110EH sold by Microfluidics, or the high pressure
homogenizer sold under the trademark ULTRA-TURRAX.RTM.). The
emulsion may be cycled through the high pressure homogenizer for
between about 2 cycles to about 100 cycles, such as about 5 cycles
to about 50 cycles or about 8 cycles to about 20 cycles (e.g.,
about any one of 8 cycles, 10 cycles, 12 cycles, 14 cycles, 16
cycles, 18 cycles or 20 cycles). The organic solvent can then be
removed by evaporation utilizing suitable equipment known for this
purpose, including, but not limited to, rotary evaporators, falling
film evaporators, wiped film evaporators, spray driers, and the
like that can be operated in batch mode or in continuous operation.
The solvent may be removed at reduced pressure (such as at about
any one of 25 mm Hg, 30 mm Hg, 40 mm Hg, 50 mm Hg, 100 mm Hg, 200
mm Hg, or 300 mm Hg). The amount of time used to remove the solvent
under reduced pressure may be adjusted based on the volume of the
formulation. For example, for a formulation produced on a 300 mL
scale, the solvent can be removed at about 1 to about 300 mm Hg
(e.g., about any one of 5-100 mm Hg, 10-50 mm Hg, 20-40 mm Hg, or
25 mm Hg) for about 5 minutes to about 60 minutes (e.g., about any
one of 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12
minutes, 13 minutes, 14 minutes, 15 minutes, 16 minutes, 18
minutes, 20 minutes, 25 minutes, or 30 minutes). The dispersion
obtained can be further lyophilized.
If desired, additional albumin solution may be added to the
dispersion to adjust the albumin to rapamycin ratio, or to adjust
the concentration of rapamycin in the dispersion. For example,
albumin solution (e.g., 25% w/v) can be added to adjust the albumin
to rapamycin ratio to about any one of 18:1, 15:1 14:1, 13:1, 12:1,
11:1, 10:1, 9:1, 8:1, 7.5:1, 7:1, 6:1, 5:1, 4:1, or 3:1. In another
example, albumin solution (e.g., 25% w/v) or another solution is
added to adjust the concentration of rapamycin in the dispersion to
about any one of 0.5 mg/ml, 1.3 mg/ml, 1.5 mg/ml, 2 mg/ml, 3 mg/ml,
4 mg/ml, 5 mg/ml, 6 mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, 10 mg/ml, 15
mg/ml, 20 mg/ml, 25 mg/ml, 30 mg/ml, 40 mg/ml, or 50 mg/ml. The
dispersion may be serially filtered through multiple filters, such
as a combination of 1.2 .mu.m and 0.8/0.2 .mu.m filters; the
combination of 1.2 .mu.m, 0.8 .mu.m, 0.45 .mu.m, and 0.22 .mu.m
filters; or the combination of any other filters known in the art.
The dispersion obtained can be further lyophilized. The
pharmaceutical compositions may be made using a batch process or a
continuous process (e.g., the production of a composition (such as
a pharmaceutical composition) on a large scale).
If desired, a second therapy (e.g., one or more compounds useful
for treating cancer), an antimicrobial agent, sugar, and/or
stabilizing agent can also be included in the composition (such as
a pharmaceutical composition). For example, this additional agent
can either be admixed with rapamycin and/or the albumin during the
preparation of the rapamycin pharmaceutical composition, or added
after the rapamycin pharmaceutical composition is prepared. In some
embodiments, the agent is admixed with the rapamycin pharmaceutical
composition prior to lyophilization. In some embodiments, the agent
is added to the lyophilized rapamycin pharmaceutical composition.
In some embodiments when the addition of the agent changes the pH
of the composition (such as a pharmaceutical composition), the pH
in the composition (such as a pharmaceutical composition) are
generally (but not necessarily) adjusted to a desired pH. Exemplary
pH values of the compositions (such as pharmaceutical compositions)
include, for example, in the range of about pH of 5 to about 8.5.
In some embodiments, the pH of the composition (such as a
pharmaceutical composition) is adjusted to no less than about pH of
6, including for example no less than any one of about pH of 6.5,
7, or 8 (e.g., about pH of 8).
Methods of Treating Diseases
Once determined suitable for medical use by following the methods
described herein, the pharmaceutical compositions may be used to
treat diseases associated with cellular proliferation or
hyperproliferation, such as cancers.
Examples of cancers that may be treated by the methods described
herein include, but are not limited to, breast cancer (such as
metastatic breast cancer), lung cancer (such as non-small cell lung
cancer), pancreatic cancer (such as metastatic pancreatic cancer or
locally advanced unresectable pancreatic cancer), multiple myeloma,
renal cell carcinoma, prostate cancer, melanoma (such as metastatic
melanoma), colon cancer, colorectal cancer, ovarian cancer, liver
cancer, renal cancer, and gastric cancer. In some embodiments, the
cancer is breast cancer after failure of combination chemotherapy
for metastatic disease or relapse within 6 months of adjuvant
chemotherapy. In some embodiments, the prior therapy includes an
anthracycline treatment.
Cancers to be treated by compositions (such as pharmaceutical
compositions) described herein include, but are not limited to,
carcinoma, lymphoma, blastoma, sarcoma, and leukemia. Examples of
cancers that can be treated by compositions (such as pharmaceutical
compositions) described herein include, but are not limited to,
squamous cell cancer, lung cancer (including small cell lung
cancer, non-small cell lung cancer, adenocarcinoma of the lung, and
squamous carcinoma of the lung, including squamous NSCLC), cancer
of the peritoneum, hepatocellular cancer, gastric or stomach cancer
(including gastrointestinal cancer), pancreatic cancer (such as
advanced pancreatic cancer), glioblastoma, cervical cancer, ovarian
cancer, liver cancer (such as hepatocellular carcinoma), bladder
cancer, hepatoma, breast cancer, colon cancer, melanoma,
endometrial or uterine carcinoma, salivary gland carcinoma, kidney
or renal cancer, prostate cancer (such as advanced prostate
cancer), vulval cancer, thyroid cancer, hepatic carcinoma, head and
neck cancer, colorectal cancer, rectal cancer, soft-tissue sarcoma,
Kaposi's sarcoma, B-cell lymphoma (including low grade/follicular
non-Hodgkin's lymphoma (NHL), small lymphocytic (SL) NHL,
intermediate grade/follicular NHL, intermediate grade diffuse NHL,
high grade immunoblastic NHL, high grade lymphoblastic NHL, high
grade small non-cleaved cell NHL, bulky disease NHL, mantle cell
lymphoma, AIDS-related lymphoma, and Waldenstrom's
macroglobulinemia), chronic lymphocytic leukemia (CLL), acute
lymphoblastic leukemia (ALL), myeloma, Hairy cell leukemia, chronic
myeloblastic leukemia, and post-transplant lymphoproliferative
disorder (PTLD), as well as abnormal vascular proliferation
associated with phakomatoses, edema (such as that associated with
brain tumors), and Meigs' syndrome. In some embodiments, there is
provided a method of treating metastatic cancer (that is, cancer
that has metastasized from the primary tumor). In some embodiments,
there is provided a method of reducing cell proliferation and/or
cell migration. In some embodiments, there is provided a method of
treating hyperplasia, for example hyperplasia in the vascular
system that can result in restenosis or hyperplasia that can result
in arterial or venous hypertension.
In some embodiments, there are provided methods of treating cancer
at advanced stage(s). In some embodiments, there are provided
methods of treating breast cancer (which may be HER2 positive or
HER2 negative), including, for example, advanced breast cancer,
stage IV breast cancer, locally advanced breast cancer, and
metastatic breast cancer. In some embodiments, the cancer is lung
cancer, including, for example, non-small cell lung cancer (NSCLC,
such as advanced NSCLC), small cell lung cancer (SCLC, such as
advanced SCLC), and advanced solid tumor malignancy in the lung. In
some embodiments, the cancer is ovarian cancer, head and neck
cancer, gastric malignancies, melanoma (including metastatic
melanoma), colorectal cancer, pancreatic cancer, and solid tumors
(such as advanced solid tumors). In some embodiments, the cancer is
any of (and in some embodiments selected from the group consisting
of) breast cancer, colorectal cancer, rectal cancer, non-small cell
lung cancer, non-Hodgkins lymphoma (NHL), renal cell cancer,
prostate cancer, liver cancer, pancreatic cancer, soft-tissue
sarcoma, Kaposi's sarcoma, carcinoid carcinoma, head and neck
cancer, melanoma, ovarian cancer, mesothelioma, gliomas,
glioblastomas, neuroblastomas, and multiple myeloma. In some
embodiments, the cancer is a solid tumor.
In some embodiments, the cancer to be treated is breast cancer,
such as metastatic breast cancer. In some embodiments, the cancer
to be treated is lung cancer, such as non-small cell lung cancer,
including advanced stage non-small cell lung cancer. In some
embodiments, the cancer to be treated is pancreatic cancer, such as
early stage pancreatic cancer or advanced or metastatic pancreatic
cancer. In some embodiments, the cancer to be treated is melanoma,
such as stage III or IV melanoma.
In some embodiments, the individual being treated for a
proliferative disease has been identified as having one or more of
the conditions described herein. Identification of the conditions
as described herein by a skilled physician is routine in the art
(e.g., via blood tests, X-rays, CT scans, endoscopy, biopsy,
angiography, CT-angiography, etc.) and may also be suspected by the
individual or others, for example, due to tumor growth, hemorrhage,
ulceration, pain, enlarged lymph nodes, cough, jaundice, swelling,
weight loss, cachexia, sweating, anemia, paraneoplastic phenomena,
thrombosis, etc. In some embodiments, the individual has been
identified as susceptible to one or more of the conditions as
described herein. The susceptibility of an individual may be based
on any one or more of a number of risk factors and/or diagnostic
approaches appreciated by the skilled artisan, including, but not
limited to, genetic profiling, family history, medical history
(e.g., appearance of related conditions), lifestyle or habits.
In some embodiments, the methods and/or compositions (such as
pharmaceutical compositions) used herein reduce the severity of one
or more symptoms associated with proliferative disease (e.g.,
cancer) by at least about any one of 10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, 90%, 95%, or 100% compared to the corresponding symptom
in the same individual prior to treatment or compared to the
corresponding symptom in other individuals not receiving the
methods and/or compositions (such as pharmaceutical
compositions).
In some embodiments, the composition (such as a pharmaceutical
composition) (such as a pharmaceutical composition) described
herein is used in combination with another administration modality
or treatment. For example, in some embodiments, the composition
(such as a pharmaceutical composition) is used in combination with
gemcitabine (for example for treating pancreatic cancer). In some
embodiments, the composition (such as a pharmaceutical composition)
is used in combination with carboplatin (for example for treating
lung cancer).
Dosing and Method of Administration
The amount of the pharmaceutical composition administered to an
individual (such as a human) may vary with the particular
composition, the method of administration, and the particular type
of recurrent cancer being treated. The amount should be sufficient
to produce a desirable beneficial effect. For example, in some
embodiments, the amount of the composition (such as a
pharmaceutical composition) is effective to result in an objective
response (such as a partial response or a complete response). In
some embodiments, the amount of pharmaceutical composition is
sufficient to result in a complete response in the individual. In
some embodiments, the amount of the composition (such as a
pharmaceutical composition) is sufficient to result in a partial
response in the individual. In some embodiments, the amount of the
composition (such as a pharmaceutical composition) administered
alone is sufficient to produce an overall response rate of more
than about any one of 40%, 50%, 60%, or 64% among a population of
individuals treated with the composition (such as a pharmaceutical
composition). Responses of an individual to the treatment of the
methods described herein can be determined, for example, based on
Response Evaluation Criteria in Solid Tumors (RECIST) or Cancer
Antigen (CA)-125 level. For example, when CA-125 is used, a
complete response can be defined as a return to a normal range
value of at least 28 days from the pretreatment value. A particle
response can be defined as a sustained over 50% reduction from the
pretreatment value.
In some embodiments, the amount of pharmaceutical composition is
sufficient to prolong progress-free survival of the individual (for
example as measured by RECIST or CA-125 changes). In some
embodiments, the amount of the pharmaceutical composition is
sufficient to prolong overall survival of the individual. In some
embodiments, the amount of the composition (such as a
pharmaceutical composition) is sufficient to produce clinical
benefit of more than about any one of 50%, 60%, 70%, or 77% among a
population of individuals treated with the composition (such as a
pharmaceutical composition).
In some embodiments, the amount of rapamycin in the composition
(such as a pharmaceutical composition) is below the level that
induces a toxicological effect (i.e., an effect above a clinically
acceptable level of toxicity) or is at a level where a potential
side effect can be controlled or tolerated when the composition
(such as a pharmaceutical composition) is administered to the
individual. In some embodiments, the amount of the composition
(such as a pharmaceutical composition) is close to a maximum
tolerated dose (MTD) of the composition (such as a pharmaceutical
composition) following the same dosing regimen. In some
embodiments, the amount of the composition (such as a
pharmaceutical composition) is more than about any one of 80%, 90%,
95%, or 98% of the MTD.
In some embodiments, the amount of rapamycin and/or composition is
an amount sufficient to decrease the size of a tumor, decrease the
number of cancer cells, or decrease the growth rate of a tumor by
at least about any one of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%,
90%, 95% or 100% compared to the corresponding tumor size, number
of cancer cells, or tumor growth rate in the same subject prior to
treatment or compared to the corresponding activity in other
subjects not receiving the treatment. Standard methods can be used
to measure the magnitude of this effect, such as in vitro assays
with purified enzyme, cell-based assays, animal models, or human
testing.
In some embodiments, the amount of rapamycin in the composition
(such as a pharmaceutical composition) is included in any one of
the following ranges: about 0.5 mg to about 5 mg, about 5 mg to
about 10 mg, about 10 mg to about 15 mg, about 15 mg to about 20
mg, about 20 mg to about 25 mg, about 20 mg to about 50 mg, about
25 mg to about 50 mg, about 50 mg to about 75 mg, about 50 mg to
about 100 mg, about 75 mg to about 100 mg, about 100 mg to about
125 mg, about 125 mg to about 150 mg, about 150 mg to about 175 mg,
about 175 mg to about 200 mg, about 200 mg to about 225 mg, about
225 mg to about 250 mg, about 250 mg to about 300 mg, about 300 mg
to about 350 mg, about 350 mg to about 400 mg, about 400 mg to
about 450 mg, or about 450 mg to about 500 mg. In some embodiments,
the amount of rapamycin in the composition (such as a
pharmaceutical composition) (e.g., a unit dosage form) is in the
range of about 5 mg to about 500 mg, such as about 30 mg to about
300 mg or about 50 mg to about 200 mg. In some embodiments, the
concentration of the rapamycin in the composition (such as a
pharmaceutical composition) is dilute (about 0.1 mg/ml) or
concentrated (about 100 mg/ml), including for example any one of
about 0.1 mg/ml to about 50 mg/ml, about 0.1 mg/ml to about 20
mg/ml, about 1 mg/ml to about 10 mg/ml, about 2 mg/ml to about 8
mg/ml, about 4 mg/ml to about 6 mg/ml, or about 5 mg/ml. In some
embodiments, the concentration of the rapamycin is at least about
any one of 0.5 mg/ml, 1.3 mg/ml, 1.5 mg/ml, 2 mg/ml, 3 mg/ml, 4
mg/ml, 5 mg/ml, 6 mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, 10 mg/ml, 15
mg/ml, 20 mg/ml, 25 mg/ml, 30 mg/ml, 40 mg/ml, or 50 mg/ml.
Exemplary doses of rapamycin in the pharmaceutical composition
include, but are not limited to, about any one of 25 mg/m.sup.2, 30
mg/m.sup.2, 50 mg/m.sup.2, 60 mg/m.sup.2, 75 mg/m.sup.2, 80
mg/m.sup.2, 90 mg/m.sup.2, 100 mg/m.sup.2, 120 mg/m.sup.2, 160
mg/m.sup.2, 175 mg/m.sup.2, 180 mg/m.sup.2, 200 mg/m.sup.2, 210
mg/m.sup.2, 220 mg/m.sup.2, 250 mg/m.sup.2, 260 mg/m.sup.2, 300
mg/m.sup.2, 350 mg/m.sup.2, 400 mg/m.sup.2, 500 mg/m.sup.2, 540
mg/m.sup.2, 750 mg/m.sup.2, 1000 mg/m.sup.2, or 1080 mg/m.sup.2 of
rapamycin. In various embodiments, the composition (such as a
pharmaceutical composition) includes less than about any one of 350
mg/m.sup.2, 300 mg/m.sup.2, 250 mg/m.sup.2, 200 mg/m.sup.2, 150
mg/m.sup.2, 120 mg/m.sup.2, 100 mg/m.sup.2, 90 mg/m.sup.2, 50
mg/m.sup.2, or 30 mg/m.sup.2 of rapamycin. In some embodiments, the
amount of rapamycin per administration is less than about any one
of 25 mg/m.sup.2, 22 mg/m.sup.2, 20 mg/m.sup.2, 18 mg/m.sup.2, 15
mg/m.sup.2, 14 mg/m.sup.2, 13 mg/m.sup.2, 12 mg/m.sup.2, 11
mg/m.sup.2, 10 mg/m.sup.2, 9 mg/m.sup.2, 8 mg/m.sup.2, 7
mg/m.sup.2, 6 mg/m.sup.2, 5 mg/m.sup.2, 4 mg/m.sup.2, 3 mg/m.sup.2,
2 mg/m.sup.2, or 1 mg/m.sup.2. In some embodiments, the dose of
rapamycin in the composition (such as a pharmaceutical composition)
is included in any one of the following ranges: about 1 mg/m.sup.2
to about 5 mg/m.sup.2, about 5 mg/m.sup.2 to about 10 mg/m.sup.2,
about 10 mg/m.sup.2 to about 25 mg/m.sup.2, about 25 mg/m.sup.2 to
about 50 mg/m.sup.2, about 50 mg/m.sup.2 to about 75 mg/m.sup.2,
about 75 mg/m.sup.2 to about 100 mg/m.sup.2, about 100 mg/m.sup.2
to about 125 mg/m.sup.2, about 125 mg/m.sup.2 to about 150
mg/m.sup.2, about 150 mg/m.sup.2 to about 175 mg/m.sup.2, about 175
mg/m.sup.2 to about 200 mg/m.sup.2, about 200 mg/m.sup.2 to about
225 mg/m.sup.2, about 225 mg/m.sup.2 to about 250 mg/m.sup.2, about
250 mg/m.sup.2 to about 300 mg/m.sup.2, about 300 mg/m.sup.2 to
about 350 mg/m.sup.2, or about 350 mg/m.sup.2 to about 400
mg/m.sup.2. Preferably, the dose of rapamycin in the composition
(such as a pharmaceutical composition) is about 5 mg/m.sup.2 to
about 300 mg/m.sup.2, such as about 100 mg/m.sup.2 to about 150
mg/m.sup.2, about 120 mg/m.sup.2, about 130 mg/m.sup.2, or about
140 mg/m.sup.2.
In some embodiments of any of the above aspects, the dose of
rapamycin in the composition (such as a pharmaceutical composition)
includes at least about any one of 1 mg/kg, 2.5 mg/kg, 3.5 mg/kg, 5
mg/kg, 6.5 mg/kg, 7.5 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg. In
various variations, the dose of rapamycin in the composition (such
as a pharmaceutical composition) includes less than about any one
of 350 mg/kg, 300 mg/kg, 250 mg/kg, 200 mg/kg, 150 mg/kg, 100
mg/kg, 50 mg/kg, 25 mg/kg, 20 mg/kg, 10 mg/kg, 7.5 mg/kg, 6.5
mg/kg, 5 mg/kg, 3.5 mg/kg, 2.5 mg/kg, 2 mg/kg, 1.5 mg/kg, or 1
mg/kg of rapamycin. In some embodiments, the dose of rapamycin in
the composition (such as a pharmaceutical composition) includes
less than about any one of 500 .mu.g/kg, 350 .mu.g/kg, 300
.mu.g/kg, 250 .mu.g/kg, 200 .mu.g/kg, 150 .mu.g/kg, 100 .mu.g/kg,
50 .mu.g/kg, 25 .mu.g/kg, 20 .mu.g/kg, 10 .mu.g/kg, 7.5 .mu.g/kg,
6.5 .mu.g/kg, 5 .mu.g/kg, 3.5 .mu.g/kg, 2.5 .mu.g/kg, 2 .mu.g/kg,
1.5 .mu.g/kg, 1 .mu.g/kg, or 0.5 .mu.g/kg of rapamycin.
Exemplary dosing frequencies include, but are not limited to, any
one of weekly without break; weekly, three out of four weeks; once
every three weeks; once every two weeks; weekly, two out of three
weeks. In some embodiments, the composition (such as a
pharmaceutical composition) is administered about once every 2
weeks, once every 3 weeks, once every 4 weeks, once every 6 weeks,
or once every 8 weeks. In some embodiments, the composition (such
as a pharmaceutical composition) is administered at least about any
one of 1.times., 2.times., 3.times., 4.times., 5.times., 6.times.,
or 7.times. (i.e., daily) a week. In some embodiments, the
intervals between each administration are less than about any one
of 6 months, 3 months, 1 month, 20 days, 15, days, 12 days, 10
days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2
days, or 1 day. In some embodiments, the intervals between each
administration are more than about any one of 1 month, 2 months, 3
months, 4 months, 5 months, 6 months, 8 months, or 12 months. In
some embodiments, there is no break in the dosing schedule. In some
embodiments, the interval between each administration is no more
than about a week.
The administration of the composition (such as a pharmaceutical
composition) can be over an extended period of time, such as from
about a month up to about seven years. In some embodiments, the
composition (such as a pharmaceutical composition) is administered
over a period of at least about any one of 2 months, 3 months, 4
months, 5 months, 6 months, 7 months, 8 months, 9 months, 10
months, 11 months, 12 months, 18 months, 24 months, 30 months, 36
months, 48 months, 60 months, 72 months, or 84 months. In some
embodiments, the composition (such as a pharmaceutical composition)
is administered over a period of at least one month, wherein the
interval between each administration is no more than about a week,
and wherein the dose of rapamycin at each administration is about
0.25 mg/m.sup.2 to about 75 mg/m.sup.2, such as about 0.25
mg/m.sup.2 to about 25 mg/m.sup.2 or about 25 mg/m.sup.2 to about
50 mg/m.sup.2.
In some embodiments, the dosage of rapamycin in a pharmaceutical
composition can be in the range of 5-400 mg/m.sup.2 when given on a
3 week schedule, or 5-250 mg/m.sup.2 when given on a weekly
schedule. For example, the amount of a rapamycin is about 60
mg/m.sup.2 to about 300 mg/m.sup.2 (e.g., about 260
mg/m.sup.2).
Other exemplary dosing schedules for the administration of the
pharmaceutical composition include, but are not limited to, any one
of 100 mg/m.sup.2, weekly, without break; 75 mg/m.sup.2 weekly, 3
out of four weeks; 100 mg/m.sup.2, weekly, 3 out of 4 weeks; 125
mg/m.sup.2, weekly, 3 out of 4 weeks; 125 mg/m.sup.2, weekly, 2 out
of 3 weeks; 130 mg/m.sup.2, weekly, without break; 175 mg/m.sup.2,
once every 2 weeks; 260 mg/m.sup.2, once every 2 weeks; 260
mg/m.sup.2, once every 3 weeks; 180-300 mg/m.sup.2, every three
weeks; 60-175 mg/m.sup.2, weekly, without break; 20-150 mg/m.sup.2
twice a week; and 150-250 mg/m.sup.2 twice a week. The dosing
frequency of the composition (such as a pharmaceutical composition)
may be adjusted over the course of the treatment based on the
judgment of the administering physician.
In some embodiments, the composition (such as a pharmaceutical
composition) is administered (e.g., intravenously) at 260 mg/m2
every three weeks. In some embodiments, the composition (such as a
pharmaceutical composition) is administered (e.g., intravenously)
at 220 mg/m.sup.2, every three weeks. In some embodiments, the
composition (such as a pharmaceutical composition) is administered
(e.g., intravenously) at 180 mg/m.sup.2, every three weeks. In some
embodiments, the composition (such as a pharmaceutical composition)
is administered (e.g., intravenously) at 200 mg/m.sup.2, every
three weeks. In some embodiments, the composition (such as a
pharmaceutical composition) is administered (e.g., intravenously)
at 130 mg/m.sup.2, every three weeks.
In some embodiments, the composition (such as a pharmaceutical
composition) is administered (e.g., intravenously) at 150
mg/m.sup.2 on days 1, 8, and 15 every 4 weeks. In some embodiments,
the composition (such as a pharmaceutical composition) is
administered (e.g., intravenously) at 125 mg/m2 on days 1, 8, and
15 every 4 weeks. In some embodiments, the composition (such as a
pharmaceutical composition) is administered (e.g., intravenously)
at 100 mg/m.sup.2 on days 1, 8, and 15 every 4 weeks. In some
embodiments, the composition (such as a pharmaceutical composition)
is administered (e.g., intravenously) at 75 mg/m2 on days 1, 8, and
15 every 4 weeks. In some embodiments, the composition (such as a
pharmaceutical composition) is administered (e.g., intravenously)
at 50 mg/m.sup.2 on days 1, 8, and 15 every 4 weeks.
The compositions (such as pharmaceutical compositions) described
herein allow infusion of the composition (such as a pharmaceutical
composition) to an individual over an infusion time that is shorter
than about 24 hours. For example, in some embodiments, the
composition (such as a pharmaceutical composition) is administered
over an infusion period of less than about any one of 24 hours, 12
hours, 8 hours, 5 hours, 3 hours, 2 hours, 1 hour, 30 minutes, 20
minutes, or 10 minutes. In some embodiments, the composition (such
as a pharmaceutical composition) is administered over an infusion
period of about 30 minutes. In some embodiments, the composition
(such as a pharmaceutical composition) is administered over an
infusion period between about 30 minutes to about 40 minutes.
In some embodiments, the present application provides a method of
treating cancer in an individual by parenterally administering to
the individual (e.g., a human) an effective amount of a composition
(such as a pharmaceutical composition) described herein. The
present application also provides a method of treating cancer in an
individual by intravenous, intra-arterial, intramuscular,
subcutaneous, inhalation, oral, intraperitoneal, nasally, or
intra-tracheal administering to the individual (e.g., a human) an
effective amount of a rapamycin pharmaceutical composition. In some
embodiments, the route of administration is intraperitoneal. In
some embodiments, the route of administration is intravenous,
intra-arterial, intramuscular, or subcutaneous. In various
variations, about 5 mg to about 500 mg, such as about 30 mg to
about 300 mg or about 50 to about 500 mg, of the rapamycin is
administered per dose. In some embodiments, the rapamycin is the
only pharmaceutically active agent for the treatment of cancer that
is contained in the composition (such as a pharmaceutical
composition).
Any of the compositions (such as pharmaceutical compositions)
described herein can be administered to an individual (such as
human) via various routes, including, for example, intravenous,
intra-arterial, intraperitoneal, intrapulmonary, oral, inhalation,
intravesicular, intramuscular, intra-tracheal, subcutaneous,
intraocular, intrathecal, transmucosal, transdermal, intratumoral,
direct injection into the blood vessel wall, intracranial, or
intra-cavity. In some embodiments, sustained continuous release
formulation of the composition (such as a pharmaceutical
composition) may be used. In one variation described herein,
nanoparticles (such as albumin nanoparticles) of the inventive
compositions (such as pharmaceutical compositions) can be
administered by any acceptable route including, but not limited to,
orally, intramuscularly, transdermally, intravenously, through an
inhaler or other air borne delivery systems and the like.
In some embodiments, the albumin-based rapamycin-containing
pharmaceutical compositions may be administered with a second
therapeutic compound and/or a second therapy. The dosing frequency
of the composition (such as a pharmaceutical composition) and the
second compound may be adjusted over the course of the treatment
based on the judgment of the administering physician. In some
embodiments, the first and second therapies are administered
simultaneously, sequentially, or concurrently. When administered
separately, the pharmaceutical composition and the second compound
can be administered at different dosing frequency or intervals. For
example, the composition (such as a pharmaceutical composition) can
be administered weekly, while a second compound can be administered
more or less frequently. In some embodiments, sustained continuous
release formulation of rapamycin-containing nanoparticle and/or
second compound may be used. Various formulations and devices for
achieving sustained release are known in the art. A combination of
the administration configurations described herein can be used.
In some embodiments, the cancer is breast cancer (for example
metastatic breast cancer), and the composition (such as a
pharmaceutical composition) is administered at 260 mg/m.sup.2 once
every three weeks.
In some embodiments, the cancer is pancreatic cancer (for example
advanced pancreatic cancer, or adenocarcinoma of the pancreas), and
the composition (such as a pharmaceutical composition) is
administered at 125 mg/m.sup.2 weekly, three out of four weeks. In
some embodiments, the cancer is pancreatic cancer (for example
advanced pancreatic cancer), and the composition (such as a
pharmaceutical composition) is administered at 125 mg/m.sup.2
weekly, three out of four weeks in combination with gemcitabine at
1000 mg/m.sup.2.
In some embodiments, the cancer is lung cancer (for example
non-small cell lung cancer), and the composition (such as a
pharmaceutical composition) is administered at 100 mg/m.sup.2
weekly. In some embodiments, the cancer is lung cancer (for example
non-small cell lung cancer), and the composition (such as a
pharmaceutical composition) is administered at 100 mg/m.sup.2
weekly, such as on Days 1, 8, 15 of each three weeks cycle, in
combination with carboplatin at AUC=6 mg min/mL once every three
weeks, such as on Day 1 of each three weeks cycle.
Metronomic Therapy Regimens
The present invention also provides metronomic therapy regimens for
any of the methods of treatment and methods of administration
described herein. Exemplary metronomic therapy regimens and
variations for the use of metronomic therapy regimens are discussed
below and disclosed in US2006/0263434 A1 (such as those described
in paragraphs [0138] to [0157] therein), which is hereby
incorporated by reference in its entirety. In some embodiments, the
pharmaceutical composition is administered over a period of at
least one month, wherein the interval between each administration
is no more than about a week, and wherein the dose of the rapamycin
at each administration is about 0.25% to about 25% of its maximum
tolerated dose following a traditional dosing regimen. In some
embodiments, the pharmaceutical composition is administered over a
period of at least two months, wherein the interval between each
administration is no more than about a week, and wherein the dose
of the rapamycin at each administration is about 1% to about 20% of
its maximum tolerated dose following a traditional dosing regimen.
In some embodiments, the dose of rapamycin per administration is
less than about any one of 25%, 24%, 23%, 22%, 20%, 18%, 5%1, 4%1,
13%1, 12%, 1%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the
maximum tolerated dose. In some embodiments, any pharmaceutical
composition is administered at least about any one of 1.times.,
2.times., 3.times., 4.times., 5.times., 6.times., or 7.times.
(i.e., daily) a week. In some embodiments, the intervals between
each administration are less than about any one of 6 months, 3
months, 1 month, 20 days, 15, days, 12 days, 10 days, 9 days, 8
days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In
some embodiments, the intervals between each administration are
more than about any one of 1 month, 2 months, 3 months, 4 months, 5
months, 6 months, 8 months, or 12 months. In some embodiments, the
composition (such as a pharmaceutical composition) is administered
over a period of at least about any one of 2 months, 3 months, 4
months, 5 months, 6 months, 7 months, 8 months, 9 months, 10
months, 11 months, 12 months, 18 months, 24 months, 30 months, 36
months, 48 months, 60 months, 72 months, or 84 months.
Exemplary Embodiments
Embodiment 1. In some embodiments, there is provided a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
polymers among the albumin on the nanoparticles, wherein a
percentage of albumin polymer among the albumin on the
nanoparticles being about 15% to about 40% (such as about 15% to
about 20%, about 20% to about 24.5%, about 24.5% to about 30%,
about 30% to about 35%, or about 35% to about 40%) is indicative of
suitability of the pharmaceutical composition for medical use.
Embodiment 2. In some embodiments, there is provided a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the percentage of albumin
monomers among the albumin on the nanoparticles, wherein a
percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%) is indicative of suitability of the
pharmaceutical composition for medical use.
Embodiment 3. In some further embodiments of embodiment 1, the
method further comprises determining the percentage of albumin
monomers among the albumin on the nanoparticles, wherein a
percentage of albumin monomers among the albumin on the
nanoparticles being about 40% to about 60% (such as about 40% to
about 55%, about 40% to about 54%, about 40% to about 53%, about
40% to about 52%, about 40% to about 50%, about 40% to about 48%,
or about 40% to about 46%) is indicative of suitability of the
pharmaceutical composition for medical use.
Embodiment 4. In some embodiments, there is provided a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the weight percentage of the
albumin in the nanoparticles, wherein a weight percentage of the
albumin in the nanoparticles being about 15% to about 30% is
indicative of suitability of the pharmaceutical composition for
medical use.
Embodiment 5. In some further embodiments of any one of embodiments
1-3, the method further comprises determining the weight percentage
of the albumin in the nanoparticles, wherein a weight percentage of
the albumin in the nanoparticles being about 15% to about 30% is
indicative of suitability of the pharmaceutical composition for
medical use.
Embodiment 6. In some embodiments, there is provided a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the weight ratio of albumin to
rapamycin in the nanoparticles, wherein an albumin to rapamycin
ratio of about 1:2 to about 1:6 in the nanoparticles is indicative
of suitability of the pharmaceutical composition for medical
use.
Embodiment 7. In some further embodiments of any one of embodiments
1-5, the method further comprises determining the weight ratio of
albumin to rapamycin in the nanoparticles, wherein an albumin to
rapamycin ratio of about 1:2 to about 1:6 in the nanoparticles is
indicative of suitability of the pharmaceutical composition for
medical use.
Embodiment 8. In some embodiments, there is provided a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the morphology of the
nanoparticles under cryo-TEM, wherein an irregular shape of the
nanoparticles is indicative of suitability of the pharmaceutical
composition for medical use.
Embodiment 9. In some further embodiments of any one of embodiments
1-8, the method further comprises determining the morphology of the
nanoparticles under cryo-TEM, wherein an irregular shape of the
nanoparticles is indicative of suitability of the pharmaceutical
composition for medical use.
Embodiment 10. In some embodiments, there is provided a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the thickness of the albumin
coating of the nanoparticles under cryo-TEM, wherein a thickness of
about 5-7 nm (such as about 6 nm) is indicative of suitability of
the pharmaceutical composition for medical use.
Embodiment 11. In some further embodiments of any one of
embodiments 1-9, the method further comprises determining the
thickness of the albumin coating of the nanoparticles under
cryo-TEM, wherein a thickness of about 5-7 nm (such as about 6 nm)
is indicative of suitability of the pharmaceutical composition for
medical use.
Embodiment 12. In some embodiments, there is provided a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the distribution of rapamycin in
a tumor tissue upon injection of the pharmaceutical composition
directly into the tumor tissue; wherein an enhanced rapamycin tumor
distribution is indicative of suitability of the pharmaceutical
composition for medical use.
Embodiment 13. In some further embodiments of any one of
embodiments 1-11, the method further comprises determining the
distribution of rapamycin in a tumor tissue upon injection of the
pharmaceutical composition directly into the tumor; wherein an
enhanced rapamycin tumor distribution is indicative of suitability
of the pharmaceutical composition for medical use.
Embodiment 14. In some embodiments, there is provided a method of
assessing suitability of a pharmaceutical composition for medical
use in a human individual, wherein the pharmaceutical composition
comprises nanoparticles comprising rapamycin coated with albumin
and a non-nanoparticle portion comprising albumin and rapamycin,
the method comprising: determining the solubility, rapamycin
crystallinity, and a rapamycin recovery following a 0.2 micron
filtration of the pharmaceutical composition, wherein a solubility
of about 50 .mu.g/ml to about 100 .mu.g/ml in a 5% human albumin
solution, a non-crystalline state of the rapamycin, and a rapamycin
recovery date of at least about 80% is indicative of suitability of
the pharmaceutical composition for medical use.
Embodiment 15. In some further embodiments of any one of
embodiments 1-13, the method further comprises determining the
solubility of the pharmaceutical composition, wherein a solubility
of about 50 .mu.g/ml to about 100 .mu.g/ml in a 5% human albumin
solution is indicative of suitability of the pharmaceutical
composition for medical use.
Embodiment 16. In some further embodiments of any one of
embodiments 1-13 and 15, the method further comprises determining
the rapamycin crystallinity of the pharmaceutical composition,
wherein a non-crystalline state of the rapamycin is indicative of
suitability of the pharmaceutical composition for medical use.
Embodiment 17. In some further embodiments of any one of
embodiments 1-13 and 15, the method further comprises determining
the rapamycin recovery following a 0.2 micron filtration of the
pharmaceutical composition, wherein a rapamycin recovery of at
least about 80% is indicative of suitability of the pharmaceutical
composition for medical use.
Embodiment 18. In some further embodiments of any one of
embodiments 15-17, the determination of solubility, rapamycin
crystalline state, or rapamycin recovery is carried out after
storage.
Embodiment 19. In some further embodiments of any one of
embodiments 14 and 16-18, the rapamycin crystallinity is determined
by X-ray diffraction, polarized light microscopy, or both.
Embodiment 20. In some further embodiments of any one of
embodiments 1-19, the method further comprises determining the
binding affinity of albumin to rapamycin in the pharmaceutical
composition.
Embodiment 21. In some further embodiments of embodiment 20, the
binding affinity is determined by equilibrium dialysis, FTIR, NMR,
or a combination thereof.
Embodiment 22. In some further embodiments of any one of
embodiments 1-21, the method further comprises determining the
surface-to-volume ratio of the nanoparticles in the pharmaceutical
composition.
Embodiment 23. In some further embodiments of any one of
embodiments 1-22, the method further comprises determining the
percentage of albumin dimers among the albumin on the
nanoparticles, wherein a percentage of about 15% to about 30% of
albumin dimers among the albumin on the nanoparticles is indicative
of the pharmaceutical composition for medical use.
Embodiment 24. In some further embodiments of any one of
embodiments 1-23, the method further comprises determining the
percentage of albumin oligomers among the albumin on the
nanoparticles, wherein a percentage of about 7% to about 15% of
albumin oligomers among the albumin on the nanoparticles is
indicative of the pharmaceutical composition for medical use.
Embodiment 25. In some further embodiments of any one of
embodiments 1-24, the method further comprises determining the
percentage of albumin monomers, dimers, oligomers, or polymers
among the total albumin in the pharmaceutical composition.
Embodiment 26. In some further embodiments of any one of
embodiments 1-3, 5, 7, 9, 11, 13, and 15-25, the percentage of
albumin monomers, dimers, oligomers, or polymers is carried out by
size-exclusion chromatography.
Embodiment 27. In some further embodiments of any one of
embodiments 1-26, the method further comprises determining the
particle size of the nanoparticles.
Embodiment 28. In some further embodiments of embodiment 27, the
particle size of the nanoparticles is determined by dynamic light
scattering.
Embodiment 29. In some further embodiments of any one of
embodiments 1-28, the method further comprises determining the
polydispersity index of the nanoparticles in the pharmaceutical
composition.
Embodiment 30. In some further embodiments of any one of
embodiments 1-29, the method further comprises determining the span
of size distribution ((Dv.sub.90-Dv.sub.10)/Dv.sub.50) of the
nanoparticles in the pharmaceutical composition.
Embodiment 31. In some further embodiments of any one of
embodiments 1-30, the method further comprises determining the
surface potential of the nanoparticles.
Embodiment 32. In some further embodiments of any one of
embodiments 1-31, the method further comprises determining the
percentage of the rapamycin in the nanoparticles among the total
rapamycin in the pharmaceutical composition.
Embodiment 33. In some further embodiments of embodiment 32, the
percentage of the rapamycin in the nanoparticles is determined by
reversed-phase HPLC.
Embodiment 34. In some further embodiments of any one of
embodiments 1-33, the method further comprises determining the
percentage of the albumin that is in the non-nanoparticle portion
among the total albumin in the pharmaceutical composition.
Embodiment 35. In some further embodiments of embodiment 34, the
percentage of the albumin is determined by size-exclusion
chromatography.
Embodiment 36. In some further embodiments of any one of
embodiments 1-35, the method further comprises determining the
stability of the pharmaceutical composition.
Embodiment 37. In some further embodiments of embodiment 36, the
stability is determined after storage.
Embodiment 38. In some further embodiments of any one of
embodiments 1-11 and 14-37, the method further comprises
determining tumor distribution of rapamycin upon administration in
vivo.
Embodiment 39. In some further embodiments of embodiment 38, the
method comprises determining tumor distribution of rapamycin upon
injection of the pharmaceutical composition directly into the tumor
tissue.
Embodiment 40. In some further embodiments of any one of
embodiments 1-39, the weight ratio of the total albumin to the
total rapamycin in the pharmaceutical composition is about 3:1 to
about 7.9:1 or about 10:1 to about 17:1.
Embodiment 41. In some further embodiments of any one of
embodiments 1-40, the albumin is human albumin.
Embodiment 42. In some further embodiments of any one of
embodiments 1-41, the average particle size of the nanoparticles is
less than about 200 nm.
Embodiment 43. In some embodiments, there is provided a method of
validating a commercial batch of a pharmaceutical composition for
medical use in a human individual, wherein the pharmaceutical
composition comprises nanoparticles comprising rapamycin coated
with albumin and a non-nanoparticle portion comprising albumin and
rapamycin, and wherein the method comprises 1) obtaining a sample
from the commercial batch, and 2) assessing suitability of the
sample for medical use according to any one of embodiments
1-42.
Embodiment 44. In some embodiments, there is provided a commercial
batch of a pharmaceutical composition for medical use in a human
individual, wherein the pharmaceutical composition comprises
nanoparticles comprising rapamycin coated with albumin and a
non-nanoparticle portion comprising albumin and rapamycin, and
wherein the commercial batch is validated by assessment of
suitability for medical use according to any one of embodiments
1-42.
EXAMPLES
The examples below are intended to be purely exemplary of the
invention and should therefore not be considered to limit the
invention in any way. The following examples and detailed
description are offered by way of illustration and not by way of
limitation.
Example 1. Determination of the Distribution of Paclitaxel within
Tumor Tissue
This example demonstrates the measurement of the distribution of
paclitaxel in tumor tissue. Distribution of paclitaxel activity was
used to monitor drug penetration and tumor cell uptake at defined
radial distances extending from a site of microinjection for three
formulations of paclitaxel.
To generate xenografts, athymic Nude-Foxn1nu mice (Jackson
Laboratories) were injected subcutaneously with 2.5.times.10.sup.6
MIA PaCa-2, A2058 melanoma, or H2122 non-small cell lung cancer
(NSCLC) cells in a 1:1 ratio with BD Biosciences Matrigel
Matrix.
Three formulations of paclitaxel were prepared for this study:
paclitaxel (PTX) solubilized in DMSO (PTX:DMSO), paclitaxel
solubilized in oil-based solvent Cremophor EL (PTX:CrEL), and a
nanoparticle formulation of paclitaxel and albumin, nab-paclitaxel
sold under the trademark ABRAXANE.RTM. (ABX). The concentration of
paclitaxel in each formulation was determined by liquid
chromatography mass spectrometry (LC-MS).
Microinjections were performed using an arrayed microinjection
device such as those sold under the trademark CIVO.RTM. (Presage
Biosciences, Seattle, Wash.) by inserting the device
transcutaneously into flank tumors of anesthetized mice. A minimum
of 3 tumors per time point were used with 2-3 replicate injection
sites per formulation in each tumor. An average drug volume of 3
.mu.L was delivered via an extrusion method over an injection
column length of 6 mm. An equal amount of paclitaxel in each
formulation (12 .mu.g) was administered per injection. Inactivated
near infrared dye, a fluorochrome sold under the trademark
VIVOTAG-S.RTM. 680 (50 .mu.g/mL), was co-injected with each drug to
mark the injection site.
Tumors were analyzed 24, 48, or 72 hours post-drug microinjection
for mitotic arrest by immunofluorescent staining of phospho-histone
H3 (pHH3). At these post-injection time points, animals were
euthanized. Tumors were harvested and resected, fixed in 10%
buffered formalin for 48 hours, and scanned on a Xenogen IVIS in
the near infrared spectrum (excitation 680 nm, emission 720 nm) to
confirm the location of each injection site. Each tumor was cut
into 2 mm thick cross sections perpendicular to the plane of
injection to enable a three-dimensional assessment of the entire
injection column.
Following IVIS imaging, tumors were processed for standard paraffin
embedding and histological analysis. 4 micron sections, cut from
each 2 mm cross section, were stained with an anti-pHH3 antibody
and a secondary antibody sold under the trademark ALEXA FLUOR
488.RTM. to assess drug-induced tumor responses (mitotic arrest)
using custom software (sold under the trademark CIVOANALYZER.TM.;
Presage Biosciences, Seattle, Wash.). Tissue sections were stained
with 4', 6-diamidino-2-phenylindole (DAPI) to visualize nuclei.
Mean fraction values of pHH3 positive cells were plotted with
standard error bars, as a function of radial distance for each
formulation and time point. To assess the statistical significance
of the differences between any pair of formulations, a linear mixed
model approach was used. In the model, the response to the PTX:CrEL
formulation was assumed to be a random effect and the differential
response due to nab-paclitaxel sold under the trademark
ABRAXANE.RTM. or PTX:DMSO was assumed to be a fixed effect. A
p-value of less than 0.05, adjusted for multiple comparisons,
indicates statistically significant differences. Data are expressed
as mean plus/minus standard error.
Immunofluorescent staining of pHH3 was used as a pharmacodynamics
indicator of paclitaxel activity to monitor drug penetration and
tumor cell uptake for MIA PaCa-2 xenografts at defined radial
distances extending from the site of injection. FIG. 1 shows
representative imaging of pancreatic MIA PaCa-2 at 24, 48, and 72
hours following microinjection with ABX, PTX:DMSO, or PTX:CrEL. At
all three time points (24, 48, and 72 hours), the area of response
and the total fraction of pHH3 positive cells at a specific radial
distance following drug microinjection were significantly greater
for nab-paclitaxel sold under the trademark ABRAXANE.RTM. compared
with either PTX:DMSO or PTX:CrEL (p<0.01)(FIG. 2A-C).
Microinjections of paclitaxel formulations were also tested on two
additional tumor xenografts, namely, A2058 melanoma and H2122 NSCLC
xenografts. Results for microinjection of A2058 xenografts showed
that ABX induced a larger increase in both the area of response and
total fraction of cells arrested in mitosis at 24 hours
post-injection when compared to PTX:CrEL (FIG. 3A)(n=5 tumors;
p<0.001). Results for microinjection of H2122 xenografts showed
that ABX induced a larger increase in both the area of response and
total fraction of cells arrested in mitosis at 24 hours
post-injection when compared to PTX:DMSO microinjection of (FIG.
3B)(n=3 tumors; p<0.001).
Example 2. Determination of the Distribution of Paclitaxel within
Tumor Tissue for Increasing Concentrations of Paclitaxel
This example demonstrates the measurement of the distribution of
paclitaxel in tumor tissue. Distribution of paclitaxel activity was
used to monitor drug penetration and tumor cell uptake at defined
radial distances extending from a site of microinjection for three
formulations of paclitaxel at three paclitaxel concentrations.
MIA PaCa-2 xenografts were generated as discussed in Example 1.
Three paclitaxel formulations were prepared for this study:
paclitaxel (PTX) solubilized in DMSO (PTX:DMSO), paclitaxel
solubilized in oil-based solvent Cremophor EL (PTX:CrEL), and a
nanoparticle formulation of paclitaxel and albumin, nab-paclitaxel
sold under the trademark ABRAXANE.RTM. (ABX). The concentration of
paclitaxel in each formulation was determined by liquid
chromatography mass spectrometry (LC-MS).
Microinjections were performed using an arrayed microinjection
device sold under the trademark CIVO.RTM. (Presage Biosciences,
Seattle, Wash.) by inserting the device transcutaneously into flank
tumors of anesthetized mice. A minimum of 3 tumors per time point
were used with 2-3 replicated injection sites per formulation in
each tumor. An average drug volume of 3 .mu.L was delivered via an
extrusion method over an injection column length of 6 mm.
Paclitaxel, as measured in PTX:DMSO, PTX:CrEL, and ABX, was
administered at three concentrations: 1.6 mg/mL, 2.5 mg/mL, or 4.75
mg/mL. Inactivated near infrared dye, sold under the trademark
VIVOTAG-S.RTM. 680 (50 .mu.g/mL), was co-injected with each drug to
mark the injection site.
Tumors were analyzed at 24 hours post-drug microinjection for
mitotic arrest by immunofluorescent staining of phospho-histone H3
(pHH3). Tumor tissue samples were prepared and analyzed as
discussed in Example 1.
Immunofluorescent staining of pHH3 was used as a pharmacodynamics
indicator of paclitaxel activity to monitor drug penetration and
tumor cell uptake at defined radial distances extending from the
site of injection for 3 concentrations of paclitaxel. At 24 hours,
the area of response and the total fraction of pHH3 positive cells
at a specific radial distance were greater for the three
concentrations of microinjected nab-paclitaxel sold under the
trademark ABRAXANE.RTM. compared with respective concentrations of
either PTX:DMSO and PTX:CrEL (FIG. 4A-C).
Example 3. Ultracentrifugation of Nanoparticles in the
Composition
This example demonstrates a method for separating nanoparticles in
the composition (such as a pharmaceutical composition) from a
non-nanoparticle portion of a composition (such as paclitaxel and
albumin). Ultracentrifugation was performed at speeds and durations
that allow the sedimentation of the nanoparticles without
significant sedimentation of any albumin and/or paclitaxel not
associated with the nanoparticles.
A pharmaceutical composition comprising nanoparticles comprising
paclitaxel coated with albumin and a non-nanoparticle portion
comprising albumin and paclitaxel was obtained. The composition was
reconstituted with 0.9% sodium chloride to yield a 5 mg/mL
suspension as measured by paclitaxel. 4.0 mL aliquots of
reconstituted suspension were transferred to quick seal polyallomer
bell-top tubes sold under the trademark BECKMAN COULTER.RTM. and
submitted to ultracentrifugation at 50,000 rpm for 41 minutes at
25.degree. C. in a Type 00 Ti rotor. After ultracentrifugation, the
tubes were removed without disturbing the pelleted nanoparticles. A
micropipette was used to remove 3.0 mL of supernatant from each
tube. Each supernatant was transferred to a separate test tube and
saved for further analysis. The remaining supernatant was poured
out of the tube containing the pelleted nanoparticles. The pellet
was then gently washed with 2 mL of water. The water was poured out
of the tube and the pellet was again gently washed with 2 mL of
water.
Example 4. Determining the Concentration of Paclitaxel and Albumin
in a Non-Nanoparticle Portion of a Composition
This example demonstrates a method for measuring the concentration
of paclitaxel and albumin from a non-nanoparticle portion of a
composition (such as a pharmaceutical composition).
As demonstrated in Example 3, the non-nanoparticle portion of a
pharmaceutical composition was separated from the nanoparticle
portion of the pharmaceutical composition by ultracentrifugation.
To determine the concentration of albumin in the non-nanoparticle
portion of the pharmaceutical composition, 1.0 mL of the
supernatant obtained following ultracentrifugation was transferred
to a 10-mL volumetric flask. The supernatant was then diluted to
10-mL with 0.9% sodium chloride. This dilute solution was then
subjected to HPLC analysis as discussed below.
Briefly, human albumin standards were created with a concentration
of 0.4 mg/mL in 0.9% sodium chloride. An HPLC system equipped with
a variable wavelength UV/VIS detector and data acquisition system
was set up with a TosoHaas TSK Guard Column SWxL (6.0 mm.times.40
mm, 7 .mu.m particle size) and a TosoHaas Analytical Column G3000
SWxL (7.8 mm.times.300 mm, 5 .mu.m particle size) kept at ambient
temperature. Separate 10 .mu.L injections of the albumin standards
or dilute supernatant samples obtained from ultracentrifugation of
the pharmaceutical composition were analyzed on the HPLC system at
228 nm using a 60-minute chromatography cycle with a flow rate of
1.0 mL/min using 100 mM K.sub.2HPO.sub.4 (pH 7.0) mobile phase.
After each chromatographic cycle, the entire line of the HPLC
system, including pump and columns, were washed with a 0.05% sodium
azide solution. The concentration of albumin in the
non-nanoparticle portion of the pharmaceutical composition was
calculated using the information obtained from the chromatograms of
the human albumin standards and the samples.
To determine the concentration of paclitaxel in the
non-nanoparticle portion of the pharmaceutical composition, 1.0 mL
of the supernatant obtained following ultracentrifugation was
transferred to a 25-mL volumetric flask. The supernatant was then
diluted to 25.0 mL with purified water, sonicated for 5 minutes,
and then allowed to cool to room temperature. 5.0 mL of this
solution was then transferred into a 10-mL volumetric flask,
diluted to 10.0 mL with a 50:50 solution of acetonitrile and water,
sonicated for 5 minutes, and then allowed to cool to room
temperature. This solution was then subjected to HPLC analysis as
discussed below.
Briefly, paclitaxel standards were created with a concentration of
1.2 mg/mL in acetonitrile. 2.0 mL of the 1.2 mg/mL paclitaxel
solution was transferred to a 100-mL volumetric flask and diluted
to volume with acetonitrile to obtain a solution of 24 .mu.g/mL
paclitaxel. 2.0 mL of the 24 .mu.g/mL paclitaxel solution was
transferred to a 25-mL volumetric flask and diluted to volume with
acetonitrile to obtain a solution of 1.9 .mu.g/mL paclitaxel. This
standard solution was stored at 4.degree. C. until use.
Additionally, a system sensitivity solution of paclitaxel was
prepared. 2.0 mL of the 1.9 .mu.g/mL paclitaxel solution was
transferred to a 50-mL volumetric flask and diluted to volume with
acetonitrile to obtain a solution of 0.08 .mu.g/mL paclitaxel. An
HPLC system equipped with a UV absorbance detector and data
acquisition system was set up with a Phenomenex, Curosil PFP guard
column (4.6 mm.times.30 mm, 5 .mu.m particle size) and a
Phenomenex, Curosil PFP analytical column (4.6 mm.times.250 mm, 5
.mu.m particle size). The autosampler was maintained at 4.degree.
C. Separate 10 .mu.L injections of the paclitaxel system
sensitivity solution, paclitaxel standard or dilute supernatant
samples obtained from ultracentrifugation of the pharmaceutical
composition were analyzed on the HPLC system at 228 nm using a
10-minute chromatography cycle with a flow rate of 1.0 mL/min using
a 70:30 acetonitrile:water mobile phase. To check the system
suitability for sample injection, the 1.9 .mu.g/mL paclitaxel
standard was injected and analyzed, followed by an injection and
analysis of 100% acetonitrile, and then an injection and analysis
of the system sensitivity solution of paclitaxel. The height of the
interference peak in the acetonitrile was confirmed to be not more
than one-fourth of the paclitaxel peak height in the analysis of
the system sensitivity solution of paclitaxel. Additionally, it was
confirmed that the signal-to-noise ratio of the paclitaxel peak in
the system sensitivity solution of paclitaxel was not less than 10.
The concentration of paclitaxel in the non-nanoparticle portion of
the pharmaceutical composition was calculated using the information
obtained from the chromatograms of the paclitaxel standard and the
samples.
Example 5. Determining the Concentration of Paclitaxel and Albumin
in a Nanoparticle Portion of a Composition
This example demonstrates a method for measuring the concentration
of paclitaxel and albumin from a nanoparticle portion of a
composition (such as a pharmaceutical composition).
As demonstrated in Example 3, the nanoparticle portion of a
pharmaceutical composition was separated from the non-nanoparticle
portion of the pharmaceutical composition by ultracentrifugation.
To determine the concentration of albumin in the nanoparticle
portion of the pharmaceutical composition, 3.0 mL of ethanol (200
absolute proof) was added to the pellet and sonicated until the
pellet was fully dispersed. After the pellet was dispersed, the
solution was transferred with a glass pipette to another centrifuge
tube. The original tube was rinsed with 2 mL ethanol and
transferred to the other centrifuge tube. The sample was then
centrifuged at 10,000 rpm for 20 minutes at 25.degree. C. After
centrifugation, the ethanol was removed with a long glass pipette.
The pellet was then dried in a desiccator under vacuum for about
1-2 hours. 3.0 mL of 0.9% sodium chloride was added to the dried
pellet and then sonicated to disperse the pellet into a homogeneous
mixture. The mixture was then transferred into a 10-mL volumetric
flask, using 5.0 mL of 0.9% sodium chloride to wash and transfer
the mixture, and then diluted to volume using 0.9% sodium chloride.
The solution was sonicated until dissolved into a homogeneous
solution and then allowed to cool to room temperature. This
solution was then subjected to HPLC analysis as discussed in
Example 4.
As demonstrated in Example 3, the nanoparticle portion of a
pharmaceutical composition was separated from the non-nanoparticle
portion of the pharmaceutical composition by ultracentrifugation.
To determine the concentration of paclitaxel in the nanoparticle
portion of the pharmaceutical composition, 3.0 mL of a 50:50
acetonitrile:water solution was added to the pellet and then
sonicated until the pellet dispersed into a homogeneous mixture.
The mixture was transferred into a 250-mL volumetric flask and
additional 50:50 acetonitrile:water solution was used to wash the
tube that contained the pellet. The solution was then diluted to
volume using the 50:50 acetonitrile:water solution, sonicated until
the pellet was completely dissolved, and then allowed to cool to
room temperature. 2.0 mL of this solution was transferred into a
100-mL volumetric flask and diluted to volume with the 50:50
acetonitrile:water solution. This solution was then subjected to
HPLC analysis as discussed in Example 4.
Example 6. Determining the Percentage of Albumin Monomers, Dimers,
Oligomers, and Polymers Among the Albumin on the Nanoparticles in
the Composition
This example demonstrates the measurement of albumin monomers,
dimers, oligomers, and polymers among the total albumin either in
the nanoparticle portion of the pharmaceutical composition or in
the non-nanoparticle portion of the pharmaceutical composition.
HPLC analysis of albumin in a nanoparticle portion of a composition
and albumin in a non-nanoparticle portion of a composition was
performed as discussed herein.
Chromatograms were generated as discussed above. For example, a
chromatogram from the method was generated as shown in FIG. 5. The
RRT, as compared to monomeric albumin, for the separated albumin
species was determined and used to classify peaks in the
chromatographs as monomeric, dimeric, oligomeric, or polymeric
albumin. The percentage of albumin monomers, dimers, oligomers, and
polymers in the nanoparticle portion of the composition was
calculated by comparing the integrated peak areas of each albumin
species to the total integrated peak area of the albumin on the
nanoparticles.
Example 7. Determining the Weight Percentage of Albumin and
Paclitaxel in the Nanoparticles in the Composition
This example demonstrates the measurement of weight percentage of
albumin on the nanoparticles and paclitaxel in the nanoparticles in
the pharmaceutical composition.
A pharmaceutical composition comprising nanoparticles comprising
paclitaxel coated with albumin and a non-nanoparticle portion
comprising albumin and paclitaxel was obtained. If necessary, the
composition was reconstituted. The nanoparticles in the composition
were then isolated away from the non-nanoparticle portion of the
composition by ultracentrifugation as discussed above.
Isolated nanoparticles were diluted and the amount of albumin and
paclitaxel in the nanoparticles was measured. The amount of albumin
on the nanoparticle is measured as detailed herein. The amount of
paclitaxel was measured by a RP-HPLC method as detailed herein.
Paclitaxel was detected at 228 nm.
The weight percentage of albumin on the nanoparticles was
determined from the amount of albumin on the nanoparticles and the
total weight of the nanoparticles (e.g., the amount of albumin and
the amount of paclitaxel). The weight ratio of albumin on the
nanoparticles to paclitaxel in the nanoparticles was determined
from the amount of albumin on the nanoparticles and the amount of
paclitaxel in the nanoparticles.
Example 8. Determining the Morphology and Thickness of the
Nanoparticles in the Composition Using Cryo-TEM
This example demonstrates a cryo-TEM technique for determining the
morphology and thickness of the nanoparticles in the pharmaceutical
composition.
A pharmaceutical composition comprising nanoparticles comprising
paclitaxel coated with albumin and a non-nanoparticle portion
comprising albumin and paclitaxel was obtained. If necessary, the
composition was reconstituted. Optionally, the nanoparticles in the
composition are then isolated away from the non-nanoparticle
portion of the composition by ultracentrifugation as discussed
above.
Nanoparticles were prepared for cryo-TEM imaging. In short, the
reconstituted nanoparticles were rapidly cooled to cryogenic
temperatures to form a vitreous form of the reconstituted
composition which was then analyzed. Briefly, the nanoparticles of
the composition were analyzed in their native structure at
magnifications ranging from 6,500.times. to 110,000.times. at
different areas in the sample. The morphology of the nanoparticles,
such as the irregularity of shape, rugosity, and surface-to-volume
ratio, was assessed from the cryo-TEM images. The thickness of the
albumin coating on the nanoparticles was also measured from the
cryo-TEM image.
Example 9. Determining the Solubility, Paclitaxel Crystallinity,
and Paclitaxel Recovery of the Composition Following 0.2 Micron
Filtration of the Composition
This example demonstrates the measurement of nanoparticle
solubility, paclitaxel crystallinity, and paclitaxel recovery of
the pharmaceutical composition following 0.2 micron filtration.
A pharmaceutical composition comprising nanoparticles comprising
paclitaxel coated with albumin and a non-nanoparticle portion
comprising albumin and paclitaxel was obtained. If necessary, the
composition was reconstituted. The composition was assessed for
nanoparticle solubility, paclitaxel crystallinity, and paclitaxel
recovery of the pharmaceutical composition following 0.2 micron
filtration immediately after reconstitution and after an
accelerated aging process. To age the composition, the
reconstituted composition was stored for 24 hours at 40.degree. C.
Compositions were filtered by passing the reconstituted composition
through a 0.2 micron filter.
Solubility of nanoparticles of the composition was determined by
performing dynamic light scattering measurements on a series of
concentrations of the composition after reconstitution. The
proportion of intact particles to free paclitaxel was a function of
the solubility of the particles. Thus, as measured by this method,
the solubility was determined as the concentration below which
particles were no longer detectable by dynamic light
scattering.
Paclitaxel crystallinity of the nanoparticles of the composition
was determined by performing an X-ray diffraction method and a
polarized light microscopy method after reconstitution. Optionally,
X-ray diffraction measurements were made on isolated nanoparticles.
Nanoparticles in the composition were optionally isolated away from
the non-nanoparticle portion of the composition by
ultracentrifugation as discussed herein. The isolated nanoparticles
were then optionally dried by lyophilization. Non-crystalline
paclitaxel in the nanoparticles will exhibit broad scattering
halos, indicative of an amorphous material (e.g., non-crystalline).
Crystalline paclitaxel in the nanoparticles will exhibit numerous
well-defined scattering peaks. Polarized light microscopy
measurements were performed on a suspension of the composition. To
determine the crystalline state of the nanoparticles of the
composition, a birefringence test was performed with an optical
microscope using polarized light.
Paclitaxel recovery following 0.2 micron filtration of the
composition was determined by performing RP-HPLC. Following 0.2
micron filtration, nanoparticles in the composition were optionally
isolated away from the non-nanoparticle portion of the composition
by ultracentrifugation as discussed herein. The amount of
paclitaxel in the nanoparticles recovered after the 0.2 micron
filtration was measured by a RP-HPLC method as discussed herein.
Paclitaxel was detected at 228 nm. The degree of paclitaxel
recovery was assessed by comparing the amount of paclitaxel in the
composition following 0.2 micron filtration to the amount of
paclitaxel in the composition prior to 0.2 micron filtration.
Example 10. Validating a Composition for Medical Use
This example demonstrates the validation of a composition for
medical use by assessing particle morphology, particle size,
surface potential, paclitaxel crystallinity, fraction of free and
bound paclitaxel or albumin, nature of the bond between paclitaxel
and albumin, dissolution profile, and oligomeric status of
albumin.
A pharmaceutical composition comprising nanoparticles comprising
paclitaxel coated with albumin and a non-nanoparticle portion
comprising albumin and paclitaxel was obtained. If necessary, the
composition was reconstituted. The composition was assessed for
sameness, compared to nab-paclitaxel sold under the trademark
ABRAXANE.RTM., with respect to particle morphology, particle size
(both D.sub.V50 and span or polydispersity index), surface
potential, paclitaxel crystallinity, fraction of free and bound
paclitaxel or albumin, nature of the bond between paclitaxel and
albumin, dissolution profile, and oligomeric status of albumin.
To assess for sameness with respect to particle morphology of
nanoparticles in the pharmaceutical composition, the morphology of
the nanoparticles was assessed.
To assess for sameness with respect to particle size and span or
polydispersity index of the nanoparticles in the pharmaceutical
composition, the reconstituted composition was subjected to dynamic
light scattering measurements. The particle size was measured as
the mean particle size (D.sub.50) of the nanoparticles in the
composition. The span or polydispersity index was measured as the
span of the size distribution, (D.sub.90-D.sub.10)/D.sub.50.
To assess for sameness with respect to the surface potential of
nanoparticles in the pharmaceutical composition, the zeta potential
of the nanoparticles was determined. The zeta potential was
measured using techniques such as microelectrophoresis,
electrophoretic light scattering, dynamic electrophoretic mobility,
or tunable resistive pulse sensing (TRPS).
To assess for sameness with respect to paclitaxel crystallinity of
nanoparticles in the pharmaceutical composition, the crystallinity
of paclitaxel in the nanoparticles of the composition was assessed
as discussed above.
To assess for sameness with respect to the fraction of free and
bound paclitaxel or albumin in the pharmaceutical composition, the
nanoparticle portion of the composition was separated from the
non-nanoparticle portion of the composition. Nanoparticles in the
composition are isolated away from the non-nanoparticle portion of
the composition by ultracentrifugation as discussed herein.
Isolated nanoparticles are diluted and the amount of albumin or
paclitaxel in the nanoparticles was measured. The amount of albumin
on the nanoparticle was measured by a HPLC size-exclusion
chromatography method. Albumin was detected at 228 nm. The amount
of paclitaxel was measured by a RP-HPLC method. Paclitaxel was
detected at 228 nm. The non-nanoparticle portion of the composition
is isolated and if the amount of albumin was determined in the
nanoparticles, then the amount of albumin was determined in the
non-nanoparticle portion of the composition. The amount of albumin
in the non-nanoparticle portion of the composition was measured by
a HPLC size-exclusion chromatography method. Albumin was detected
at 228 nm. If the amount of paclitaxel was determined in the
nanoparticles, then the amount of paclitaxel was determined in the
non-nanoparticle portion of the composition. The amount of
paclitaxel was measured by a RP-HPLC method. Paclitaxel was
detected at 228 nm. The fraction of free (non-nanoparticle portion)
and bound paclitaxel (nanoparticle portion) or albumin was
calculated from the measured values.
To assess for sameness with respect to the nature of the bond
between paclitaxel and albumin, the nature of the bond between
albumin and paclitaxel was assessed.
To assess for sameness with respect to the dissolution profile, the
reconstituted composition was subjected to an in vitro release
kinetics assay. The reconstituted composition was diluted in a 0.9%
saline solution and dynamic light scattering was used to monitor
particle size over 60 minutes for a range of concentrations of the
composition, or the reconstituted composition was diluted in a 5%
human albumin solution and dynamic light scattering was used to
monitor scattering and particle size over 60 minutes for a range of
concentrations of the composition, or the reconstituted composition
was diluted in water and a UV-Vis spectrophotometer was used to
monitor absorbance over 60 minutes for a range of concentrations of
the composition.
To assess for sameness with respect to the oligomeric status of
albumin, albumins in the starting material and in the final
composition were assessed for oligomeric status. Isolated albumin
in the starting material and final composition were analyzed by a
HPLC size-exclusion chromatography method. Albumin was detected at
228 nm. A chromatogram of the HPLC-size exclusion chromatography
method was generated as shown in FIG. 5. The RRT, as compared to
monomeric albumin, for the separated albumin species was determined
and used to classify peaks in the chromatographs as monomeric,
dimer, oligomeric, or polymeric albumin.
The composition was suitable for medical use if the particle
morphology, particle size, surface potential, paclitaxel
crystallinity, fraction of free and bound paclitaxel or albumin,
nature of the bond between paclitaxel and albumin, dissolution
profile, and oligomeric status of albumin were found to exhibit
sameness with respect to results obtained from analysis of
nab-paclitaxel sold under the trademark ABRAXANE.RTM..
Example 11. Validating a Composition for Medical Use
This example demonstrates the validation of a composition for
medical use, both before and after storage, by assessing particle
morphology, particle size, surface potential, paclitaxel
crystallinity, fraction of free and bound paclitaxel or albumin,
nature of the bond between paclitaxel and albumin, dissolution
profile, oligomeric status of albumin, oligomeric status of albumin
on the nanoparticle, and the recovery of paclitaxel following 0.2
micron filtration.
A pharmaceutical composition comprising nanoparticles comprising
paclitaxel coated with albumin and a non-nanoparticle portion
comprising albumin and paclitaxel was obtained. If necessary, the
composition was reconstituted. The composition was assessed for
sameness, in view of nab-paclitaxel sold under the trademark
ABRAXANE.RTM., with respect to particle morphology, particle size,
surface potential, paclitaxel crystallinity, fraction of free and
bound paclitaxel or albumin, nature of the bond between paclitaxel
and albumin, dissolution profile, oligomeric status of albumin,
oligomeric status of albumin on the nanoparticle, and the recovery
of paclitaxel following 0.2 micron filtration. To test the
composition following storage conditions, the composition was
stored for 24 hours at 40.degree. C. to simulate an accelerated
aging process.
To assess for sameness with respect to particle morphology of
nanoparticles in the pharmaceutical composition, the nanoparticles
in the composition were assessed as discussed herein. The
morphology of the nanoparticles, such as the irregularity of shape,
was assessed from the cryo-TEM images.
To assess for sameness with respect to particle size and span or
polydispersity index of the nanoparticles in the pharmaceutical
composition, the reconstituted composition was subjected to dynamic
light scattering measurements. The particle size was measured as
the volume-weighted mean particle size (D.sub.v50) of the
nanoparticles in the composition. The span or polydispersity index
was measured as the span of the volume-weighted size distribution,
(D.sub.v90-D.sub.v10)/D.sub.v50.
To assess for sameness with respect to the surface potential of
nanoparticles in the pharmaceutical composition, the zeta potential
of the nanoparticles was determined. The zeta potential was
measured using techniques such as microelectrophoresis,
electrophoretic light scattering, dynamic electrophoretic mobility,
or tunable resistive pulse sensing (TRPS).
To assess for sameness with respect to paclitaxel crystallinity of
nanoparticles in the pharmaceutical composition, paclitaxel
crystallinity of the nanoparticles of the composition was
determined by performing an X-ray diffraction method and a
polarized light microscopy method as detailed herein.
To assess for sameness with respect to the fraction of free (i.e.,
not associated with the nanoparticles) and bound (i.e., associated
with the nanoparticles) paclitaxel and albumin in the
pharmaceutical composition, the composition was assessed as
detailed herein.
To assess for sameness with respect to the nature of the bond
between paclitaxel and albumin, the composition was assessed via
equilibrium dialysis test and FTIR and NMR analysis. The nature of
the bond between paclitaxel and albumin was assessed using
paclitaxel and processed albumin. The binding affinity between
paclitaxel and processed albumin in the composition was measured
via an equilibrium dialysis testing apparatus and further by FTIR
and NMR analysis.
To assess for sameness with respect to the dissolution profile, the
reconstituted composition was subject to an in vitro release
kinetics assay. The reconstituted composition was diluted in a 0.9%
saline solution and dynamic light scattering is used to monitor
particle size over 60 minutes for a range of concentrations of the
composition, or the reconstituted composition was diluted in a 5%
human albumin solution and dynamic light scattering was used to
monitor scattering and particle size over 60 minutes for a range of
concentrations of the composition, or the reconstituted composition
was diluted in water and a UV-Vis spectrophotometer was used to
monitor absorbance over 60 minutes for a range of concentrations of
the composition.
To assess for sameness with respect to the oligomeric status of
albumin, albumins in the starting material and in the final
composition are isolated away from the other components of the
composition and their oligomeric status was assessed. Isolated
albumins in the starting product of final composition are analyzed
by a HPLC size-exclusion chromatography method as detailed
herein.
To assess for sameness with respect to the oligomeric status of
albumin on the nanoparticle, the nanoparticles in the composition
were isolated away from the non-nanoparticle portion of the
composition by ultracentrifugation as discussed herein. Isolated
nanoparticles are diluted and then analyzed by a HPLC
size-exclusion chromatography method as discussed herein.
To assess for sameness with respect to the recovery of paclitaxel
following 0.2 micron filtration, the reconstituted composition was
filtered with a 0.2 micron filter. Following 0.2 micron filtration,
nanoparticles in the composition are isolated away from the
non-nanoparticle portion of the composition by ultracentrifugation
as discussed herein. The amount of paclitaxel in the recovered
nanoparticles from the 0.2 micron filtered composition was measured
by a RP-HPLC method as discussed herein. Paclitaxel was detected at
228 nm. The degree of paclitaxel recovery was assessed by comparing
the amount of paclitaxel in the composition following 0.2 micron
filtration to the amount of paclitaxel in the composition prior to
0.2 micron filtration.
The composition was suitable for medical use if the particle
morphology, particle size, surface potential, paclitaxel
crystallinity, fraction of free and bound paclitaxel or albumin,
nature of the bond between paclitaxel and albumin, dissolution
profile, oligomeric status of albumin, status of albumin on the
nanoparticle, and the recovery of paclitaxel following 0.2 micron
filtration were found to exhibit sameness with respect to results
obtained from analysis of nab-paclitaxel sold under the trademark
ABRAXANE.RTM..
Example 12. Determining In Vitro Dissolution Kinetics of the
Pharmaceutical Composition Using a UV-Vis Spectrophotometer
This example demonstrates the measurement of in vitro release
kinetics of the pharmaceutical composition after reconstitution, as
determined by a UV-Vis spectrophotometer.
The pharmaceutical composition was reconstituted with a 0.9% sodium
chloride solution to produce a 5 mg/ml stock solution of the
pharmaceutical composition, as measured by the concentration of
paclitaxel. For example, starting from a vial of the lyophilized
pharmaceutical composition containing 100 mg, as measured by the
amount of paclitaxel, 20 ml of 0.9% sodium chloride was slowly
injected into the vial over a minimum of 1 minute using a sterile
syringe. The flow of 0.9% sodium chloride was directed onto the
inside wall of the vial. Subsequently, the lyophilized
pharmaceutical composition was allowed to rest for 5 minutes and
then the vial was gently swirled or slowly inverted for at least 2
minutes until complete dissolution of the pharmaceutical
composition occurred. The lyophilized pharmaceutical composition
was reconstituted in a manner to avoid the formation of foam. The
spectrophotometer was set up as shown in FIG. 6. A 295 nm longwave
pass filter was placed between the UV light source and the cuvette.
A 10-mm quartz cuvette was placed into an Agilent Cary 8454 UV-Vis
spectrophotometer pre-equilibrated to 20.degree. C. while stirring
at 2000 rpm. An appropriate volume of water was transferred to the
cuvette. A magnetic stirrer bar was added to the bottom of cuvette.
The spectrophotometer was equilibrated over an hour (or longer)
until the 340 nm signal was stabilized. An appropriate volume of
the stock reconstituted pharmaceutical composition suspension was
then added to the cuvette to achieve a target paclitaxel
concentration of 100 .mu.g/ml. The suspension was immediately mixed
and the cuvette was capped.
During the preparation of the 100 .mu.g/ml pharmaceutical
composition, a series of consecutive intensity measurement was
performed. All kinetic measurements were performed at 20.degree. C.
with the following settings: path length: 1 cm; wavelength range:
190-1100 nm; integration time: 0.5 seconds; interval: 1 nm;
deuterium lamp (UV): On; run time: 3600 seconds; cycle time: 0.5
seconds; wavelength: 340 nm; and stirrer speed: 2000 rpm.
As shown in FIG. 7, the majority of particles in the reconstituted
pharmaceutical composition suspension diluted to concentrations
above the paclitaxel solubility in the release media dissolved and
disintegrated rapidly, first over 30 seconds, followed by a long
slow transient release over 3600 seconds or longer. The rate of
disintegration and dissolution, and the dissolution profile shapes
were concentration dependent.
Example 13. Methods of Making a Nanoparticle Composition
This example demonstrates methods of making an albumin/paclitaxel
nanoparticle composition (such as a pharmaceutical composition) and
variants thereof.
All variants were manufactured using laboratory scale/bench-top
equipment.
Variant 5 (V5) was prepared using the following procedure.
5% human albumin (HA) solution and paclitaxel solvent solution
containing approximately 200 mg/mL paclitaxel in 90:10 v:v
chloroform:ethanol mixture were prepared. 18.4 mL of the 5% HA
solution was transferred to a beaker and mixed using a high-shear
mixer sold under the trademark SILVERSON.RTM.. 1.6 mL of the
paclitaxel solvent solution was slowly (drop-wise) added to the HA
solution and mixed for approximately 5 minutes at 5000-6000 rpm to
create a crude emulsion. The crude emulsion was high-pressure
homogenized at pressure of approximately 18-20 kpsi using an
emulsifier sold under the trademarks AVESTIN.RTM. EMULSIFLEX.TM.-C5
for approximately 12 passes to create a fine emulsion. The fine
emulsion was transferred to a 2 L round bottom flask and the
solvents were removed using a rotary evaporator sold under the
trademark BUCHI.RTM. with a water bath temperature set at
40.degree. C. The evaporation was performed using the parameters in
Table 1. The evaporation continued until the initial volume was
reduced by approximately 45-80%. The whole process was repeated
once to generate enough material for in-process (IP) and finished
product (FP) testing. The post-evaporated (PE) suspension was
combined, mixed and assayed for paclitaxel and HA. Based on the
assay values, the PE suspension was diluted with water sold under
the trademark MILLI-Q.RTM. and 20-25% HA solution to paclitaxel
concentration of 7 mg/mL and HA concentration of 56 mg/mL. The
diluted suspension was filtered through a series of 1.2 .mu.m, 0.8
.mu.m, 0.45 .mu.m and 0.2 .mu.m syringe filters with
polyethersulfone (PES) membrane sold under the trademark
SUPOR.RTM.. 1-3 mL aliquots of the filtered suspension were filled
in 10 mL, 20 mm glass vials and lyophilized in VirTis Genesis EL25
lyophilizer (SP Industries, Gardiner, N.Y.) using the cycle in
Table 2. After lyophilization the vials were stoppered under
nitrogen, crimped and stored at -20.degree. C. for future
testing.
A total of three lots of Variant 5 were manufactured.
TABLE-US-00001 TABLE 1 Evaporation cycle. Step # Pressure set point
Hold time after pressure is achieved 1 70 mm Hg 1 min 2 60 mm Hg 1
min 3 50 mm Hg 1 min 4 40 mm Hg 1 min 5 30 mm Hg 1 min 6 25 mm Hg
As needed
TABLE-US-00002 TABLE 2 Lyophilization cycle. Step Temp. (.degree.
C.) Time (min) Vac (mTorr) Type Loading 1 -55 N/A N/A Hold Freezing
1 -55 240 N/A Hold Drying 1 -55 10 350 Hold 2 -15 200 350 Ramp 3
-15 10 350 Hold 4 25 400 350 Ramp 5 25 840 350 Hold 6 30 50 350
Ramp 7 30 480 350 Hold
Variant 1 (V1) was prepared using the procedure for Variant 5 with
the following modification. The organic solvent mixture used to
prepare the 200 mg/mL paclitaxel solvent solution contained 50% by
volume ethanol and 50% by volume chloroform.
A total of three lots of Variant 1 were manufactured.
Variant 2 (V2) was prepared using the procedure for Variant 5 with
the following modification. The concentration of the human albumin
solution used to prepare the crude emulsion was 10 mg/mL.
A total of three lots of Variant 2 were manufactured.
Variant 3 (V3) was prepared using the procedure for Variant 5 with
the following modification. The high-pressure homogenization of the
crude emulsion was performed at pressure of approximately 5
kpsi.
A total of three lots of Variant 3 were manufactured.
Variant 4 (V4) was prepared using the procedure for Variant 5 with
the following modifications. After the high-pressure homogenization
the fine emulsion was transferred to a 500 mL round bottom flask
and the solvents were removed using a rotary evaporator sold under
the trademark BUCHI.RTM. with a water bath temperature set at
30.degree. C. The evaporation was performed using the parameters in
Table 3.
TABLE-US-00003 TABLE 3 Evaporation cycle for variant 4. Step #
Pressure set point Hold time after pressure is achieved 1 70 mm Hg
1 min 2 60 mm Hg 1 min 3 50 mm Hg 1 min 4 From 40 mm Hg to 25 mm 1
min Hg, every 1 mm Hg 5 15 mm Hg As needed
A total of three lots of Variant 4 were manufactured.
Example 14. Assessment of Compositions
This example reports results from the assessment of various
albumin/paclitaxel nanoparticle compositions.
The compositions were assessed using the methods described herein.
Variants are designated as discussed herein (e.g., FP refers to
finished product; IP refers to in-process product). Nab-paclitaxel
sold under the trademark ABRAXANE.RTM. is the proprietary
albumin/paclitaxel nanoparticle product of Celgene/Abraxis.
Paclitaxel NAB, Albupax, and PacliALL are purported copies of
nab-paclitaxel sold under the trademark ABRAXANE.RTM. made by
different companies.
TABLE-US-00004 TABLE 4 D.sub.V4,3 (nm) measured immediately after
reconstitution. N Margin Sample Name (number) Mean of error Min Max
ABRAXANE .RTM. 22 153.8 2.8 144.1 164.4 Paclitax NAB 3 113.7 12.7
108.5 118.7 Albupax 5 140.0 12.0 129.6 151.6 PacliALL 5 426.3 864.0
221.7 827.9 V5 FP 3 159.5 19.8 153.4 168.5 V1 FP 3 177.6 74.4 158.8
212.1 V2 FP 3 179.5 27.1 167.0 186.9 V3 FP 3 178.9 17.8 171.5 185.8
V4 FP 3 147.2 9.8 142.7 150.0
TABLE-US-00005 TABLE 5 Z average (nm) measured immediately after
reconstitution. N Margin Sample Name (number) Mean of error Min Max
ABRAXANE .RTM. 22 145.9 1.9 138.5 153.0 Paclitax NAB 3 118.7 8.5
115.2 122.0 Albupax 5 138.9 1.7 137.2 140.5 PacliALL 5 178.6 13.6
165.8 190.0 V5 FP 3 149.7 12.2 146.5 155.4 V1 FP 3 164.8 55.7 149.3
190.5 V2 FP 3 161.4 12.9 155.6 165.6 V3 FP 3 162.6 6.8 159.9 165.4
V4 FP 3 142.9 6.7 139.9 145.1
TABLE-US-00006 TABLE 6 Polydispersity index (PDI) measured
immediately after reconstitution. N Margin Sample Name (number)
Mean of error Min Max ABRAXANE .RTM. 22 0.119 0.006 0.097 0.145
Paclitax NAB 3 0.099 0.039 0.087 0.117 Albupax 5 0.117 0.011 0.109
0.132 PacliALL 5 0.166 0.060 0.118 0.241 V5 FP 3 0.127 0.027 0.118
0.139 V1 FP 3 0.092 0.079 0.071 0.129 V2 FP 3 0.128 0.059 0.109
0.155 V3 FP 3 0.111 0.041 0.093 0.126 V4 FP 3 0.098 0.024 0.088
0.107
TABLE-US-00007 TABLE 7 D.sub.V5 (nm) measured immediately after
reconstitution. N Margin Sample Name (number) Mean of error Min Max
ABRAXANE .RTM. 22 75.0 2.3 68.2 88.0 Paclitax NAB 3 62.1 12.0 59.3
67.7 Albupax 5 73.4 19.8 55.2 91.8 PacliALL 5 86.7 6.1 82.1 93.6 V5
FP 3 71.7 10.1 67.7 75.8 V1 FP 3 91.1 53.4 71.4 114.0 V2 FP 3 80.7
18.1 72.3 85.2 V3 FP 3 85.6 3.7 84.2 87.2 V4 FP 3 76.2 8.9 73.3
80.2
TABLE-US-00008 TABLE 8 D.sub.V50 (nm) measured immediately after
reconstitution. N Margin Sample Name (number) Mean of error Min Max
ABRAXANE .RTM. 22 137.9 2.6 129 148 Paclitax NAB 3 102.8 13.8 98.3
109 Albupax 5 130.6 15.3 117 146 PacliALL 5 192.0 25.6 168 214 V5
FP 3 140.7 22.3 135 151 V1 FP 3 164.0 83.1 139 202 V2 FP 3 160.7
17.4 154 168 V3 FP 3 163.3 10.0 159 167 V4 FP 3 134.3 10.0 130
138
TABLE-US-00009 TABLE 9 D.sub.V95 (nm) measured immediately after
reconstitution. N Margin Sample Name (number) Mean of error Min Max
ABRAXANE .RTM. 22 288.4 10.1 237 341 Paclitax NAB 3 207.3 23.6 198
217 Albupax 5 243.2 8.5 235 251 PacliALL 5 1263.0 2393.5 333 4710
V5 FP 3 314.0 33.4 299 325 V1 FP 3 315.3 93.2 277 352 V2 FP 3 344.0
123.0 294 393 V3 FP 3 328.0 66.1 300 353 V4 FP 3 264.3 22.5 256
274
TABLE-US-00010 TABLE 10 (D.sub.V90 - D.sub.V10)/D.sub.V50 measured
immediately after reconstitution. N Margin Sample Name (number)
Mean of error Min Max ABRAXANE .RTM. 22 1.21 0.06 0.82 1.53
Paclitax NAB 3 1.08 0.25 0.98 1.18 Albupax 5 1.05 0.30 0.79 1.36
PacliALL 5 4.35 8.40 1.11 16.46 V5 FP 3 1.35 0.20 1.30 1.45 V1 FP 3
1.10 0.60 0.94 1.38 V2 FP 3 1.30 0.63 1.08 1.58 V3 FP 3 1.17 0.23
1.07 1.26 V4 FP 3 1.10 0.12 1.05 1.15
TABLE-US-00011 TABLE 11 Paclitaxel in solution phase
(non-nanoparticle portion) as a fraction of total paclitaxel,
expressed as a percentage, immediately after reconstitution. N
Margin Sample Name (number) Mean of error Min Max ABRAXANE .RTM. 65
1.71 0.027 1.46 2.14 Paclitax NAB 1 2.64 -- 2.64 2.64 Albupax 1
1.42 -- 1.42 1.42 PacliALL 4 2.24 0.706 1.65 2.64 V5 FP 1 1.55 --
1.55 1.55 V1 FP 1 1.51 -- 1.51 1.51 V2 FP 1 1.63 -- 1.63 1.63 V3 FP
1 1.55 -- 1.55 1.55 V4 FP 1 1.59 -- 1.59 1.59 V5 IP 1 0.85 -- 0.85
0.85 V1 IP 1 0.93 -- 0.93 0.93 V2 IP 1 0.69 -- 0.69 0.69 V3 IP 1
0.71 -- 0.71 0.71 V4 IP 1 1.23 -- 1.23 1.23
TABLE-US-00012 TABLE 12 Paclitaxel in particles (nanoparticle
portion) as a fraction of total paclitaxel, expressed as a
percentage, immediately after reconstitution. N Margin Sample Name
(number) Mean of error Min Max ABRAXANE .RTM. 65 98.290 0.025
97.860 98.540 Paclitax NAB 1 97.351 -- 97.351 97.351 Albupax 1
98.575 -- 98.575 98.575 PacliALL 4 97.760 0.705 97.360 98.350 V5 FP
1 98.441 -- 98.441 98.441 V1 FP 1 98.482 -- 98.482 98.482 V2 FP 1
98.365 -- 98.365 98.365 V3 FP 1 98.442 -- 98.442 98.442 V4 FP 1
98.409 -- 98.409 98.409 V5 IP 1 99.1 -- 99.1 99.1 V1 IP 1 99.1 --
99.1 99.1 V2 IP 1 99.3 -- 99.3 99.3 V3 IP 1 99.3 -- 99.3 99.3 V4 IP
1 98.8 -- 98.8 98.8
TABLE-US-00013 TABLE 13 Albumin in solution phase (non-nanoparticle
portion) as a fraction of total albumin, expressed as a percentage,
immediately after reconstitution. N Margin Sample Name (number)
Mean of error Min Max ABRAXANE .RTM. 66 96.150 0.135 95.330 97.760
Paclitax NAB 1 98.701 -- 98.701 98.701 Albupax 1 96.648 -- 96.648
96.648 PacliALL 4 98.450 0.175 98.360 98.600 V5 FP 1 97.094 --
97.094 97.094 V1 FP 1 98.033 -- 98.033 98.033 V2 FP 1 97.961 --
97.961 97.961 V3 FP 1 96.846 -- 96.846 96.846 V4 FP 1 96.033 --
96.033 96.033 V5 IP 1 94.1 -- 94.1 94.1 V1 IP 1 96.9 -- 96.9 96.9
V2 IP 1 71.0 -- 71.0 71.0 V3 IP 1 90.8 -- 90.8 90.8 V4 IP 1 95.6 --
95.6 95.6
TABLE-US-00014 TABLE 14 Albumin in particles (nanoparticle portion)
as a fraction of total albumin, expressed as a percentage,
immediately after reconstitution. N Margin Sample Name (number)
Mean of error Min Max ABRAXANE .RTM. 66 3.846 0.134 2.240 4.670
Paclitax NAB 1 1.299 -- 1.299 1.299 Albupax 1 3.352 -- 3.352 3.352
PacliALL 4 1.555 0.173 1.400 1.640 V5 FP 1 2.906 -- 2.906 2.906 V1
FP 1 1.967 -- 1.967 1.967 V2 FP 1 2.039 -- 2.039 2.039 V3 FP 1
3.154 -- 3.154 3.154 V4 FP 1 3.967 -- 3.967 3.967 V5 IP 1 5.9 --
5.9 5.9 V1 IP 1 3.1 -- 3.1 3.1 V2 IP 1 29.0 -- 29.0 29.0 V3 IP 1
9.2 -- 9.2 9.2 V4 IP 1 4.4 -- 4.4 4.4
TABLE-US-00015 TABLE 15 Concentration of free (in solution phase)
paclitaxel (.mu.g/ml) in the composition immediately after
reconstitution. Sample Name N (number) Mean Margin of error Min Max
ABRAXANE .RTM. 65 80.6 1.2 68.4 94.2 Paclitax NAB 1 125.0 -- 125.0
125.0 Albupax 1 71.5 -- 71.5 71.5 PacliALL 4 101.1 30.6 73.9 116.1
V5 FP 1 75.0 -- 75.0 75.0 V1 FP 1 70.4 -- 70.4 70.4 V2 FP 1 71.6 --
71.6 71.6 V3 FP 1 70.6 -- 70.6 70.6 V4 FP 1 77.5 -- 77.5 77.5 V5 IP
1 130.4 -- 130.4 130.4 V1 IP 1 115.8 -- 115.8 115.8 V2 IP 1 71.2 --
71.2 71.2 V3 IP 1 102.0 -- 102.0 102.0 V4 IP 1 125.0 -- 125.0
125.0
TABLE-US-00016 TABLE 16 Concentration of bound (in
particles/nanoparticle portion) paclitaxel (.mu.g/ml) in the
composition immediately after reconstitution. Margin of Sample Name
N (number) Mean error Min Max ABRAXANE .RTM. 66 4624 45 4156 5147
Paclitax NAB 1 4594 -- 4594 4594 Albupax 1 4945 -- 4945 4945
PacliALL 4 4405 176 4281 4547 V5 FP 1 4737 -- 4737 4737 V1 FP 1
4568 -- 4568 4568 V2 FP 1 4307 -- 4307 4307 V3 FP 1 4461 -- 4461
4461 V4 FP 1 4795 -- 4795 4795 V5 IP 1 15058.0 -- 15058.0 15058.0
V1 IP 1 12282.4 -- 12282.4 12282.4 V2 IP 1 10164.6 -- 10164.6
10164.6 V3 IP 1 14152.6 -- 14152.6 14152.6 V4 IP 1 9967.2 -- 9967.2
9967.2
TABLE-US-00017 TABLE 17 Concentration of free (in solution phase)
albumin (mg/ml) in the composition immediately after
reconstitution. Sample Name N (number) Mean Margin of error Min Max
ABRAXANE .RTM. 66 39.0 0.485 34.9 46.4 Paclitax NAB 1 60.8 -- 60.8
60.8 Albupax 1 34.4 -- 34.4 34.4 PacliALL 4 42.7 1.005 41.8 43.3 V5
FP 1 40.1 -- 40.1 40.1 V1 FP 1 36.6 -- 36.6 36.6 V2 FP 1 44.5 --
44.5 44.5 V3 FP 1 36.9 -- 36.9 36.9 V4 FP 1 37.4 -- 37.4 37.4 V5 IP
1 51.4 -- 51.4 51.4 V1 IP 1 47.8 -- 47.8 47.8 V2 IP 1 7.3 -- 7.3
7.3 V3 IP 1 52.9 -- 52.9 52.9 V4 IP 1 51.5 -- 51.5 51.5
TABLE-US-00018 TABLE 18 Concentration of bound (in
particles/nanoparticle portion) albumin (mg/ml) in the composition
immediately after reconstitution. Sample Name N (number) Mean
Margin of error Min Max ABRAXANE .RTM. 66 1.555 0.052 0.800 1.900
Paclitax NAB 1 0.800 -- 0.800 0.800 Albupax 1 1.193 -- 1.193 1.193
PacliALL 4 0.676 0.087 0.595 0.715 V5 FP 1 1.199 -- 1.199 1.199 V1
FP 1 0.735 -- 0.735 0.735 V2 FP 1 0.926 -- 0.926 0.926 V3 FP 1
1.201 -- 1.201 1.201 V4 FP 1 1.544 -- 1.544 1.544 V5 IP 1 3.234 --
3.234 3.234 V1 IP 1 1.520 -- 1.520 1.520 V2 IP 1 2.996 -- 2.996
2.996 V3 IP 1 5.376 -- 5.376 5.376 V4 IP 1 2.382 -- 2.382 2.382
TABLE-US-00019 TABLE 19 Albumin as a percentage of the nanoparticle
mass immediately after reconstitution. Sample Name N (number) Mean
Margin of error Min Max ABRAXANE .RTM. 30 24.0 1.4 13.6 29.0
Paclitax NAB 1 14.8 -- 14.8 14.8 Albupax 1 19.4 -- 19.4 19.4
PacliALL 4 13.3 1.7 11.9 14.3 V5 FP 1 20.2 -- 20.2 20.2 V1 FP 1
13.9 -- 13.9 13.9 V2 FP 1 17.7 -- 17.7 17.7 V3 FP 1 21.2 -- 21.2
21.2 V4 FP 1 24.4 -- 24.4 24.4 V5 IP 1 17.7 -- 17.7 17.7 V1 IP 1
11.0 -- 11.0 11.0 V2 IP 1 22.8 -- 22.8 22.8 V3 IP 1 27.5 -- 27.5
27.5 V4 IP 1 19.3 -- 19.3 19.3
A comparison of the albumin as a percentage of the nanoparticle
mass (Table 19) is illustrated in the bar graph of FIG. 8. Amongst
all the compositions, nab-paclitaxel sold under the trademark
ABRAXANE.RTM. and Paclitax NAB are most stable.
TABLE-US-00020 TABLE 20 Paclitaxel as a percentage of the
nanoparticle mass immediately after reconstitution. Sample Name N
(number) Mean Margin of error Min Max ABRAXANE .RTM. 30 76.1 1.435
71.0 86.4 Paclitax NAB 1 85.2 -- 85.2 85.2 Albupax 1 80.6 -- 80.6
80.6 PacliALL 4 86.7 1.695 85.7 88.1 V5 FP 1 79.8 -- 79.8 79.8 V1
FP 1 86.1 -- 86.1 86.1 V2 FP 1 82.3 -- 82.3 82.3 V3 FP 1 78.8 --
78.8 78.8 V4 FP 1 75.6 -- 75.6 75.6 V5 IP 1 82 -- 82 82 V1 IP 1 89
-- 89 89 V2 IP 1 77 -- 77 77 V3 IP 1 72 -- 72 72 V4 IP 1 81 -- 81
81
TABLE-US-00021 TABLE 21 Percentage of albumin in the form of
monomers on the nanoparticles. Sample Name N (number) Mean Margin
of error Min Max ABRAXANE .RTM. 66 47.1 2.5 20.3 66.9 Paclitax NAB
1 44.8 -- 44.8 44.8 Albupax 1 55.6 -- 55.6 55.6 PacliALL 4 54.1
20.2 40.7 70.9 V5 FP 1 68.5 -- 68.5 68.5 V1 FP 1 72.8 -- 72.8 72.8
V2 FP 1 61.9 -- 61.9 61.9 V3 FP 1 56.3 -- 56.3 56.3 V4 FP 1 63.5 --
63.5 63.5 V5 IP 1 66.1 -- 66.1 66.1 V1 IP 1 78.1 -- 78.1 78.1 V2 IP
1 51.9 -- 51.9 51.9 V3 IP 1 55.7 -- 55.7 55.7 V4 IP 1 69.9 -- 69.9
69.9
A comparison of the percentage of albumin on the nanoparticles in
the form of monomers (Table 21) is illustrated in the bar graph of
FIG. 9. Amongst all the compositions, nab-paclitaxel sold under the
trademark ABRAXANE.RTM. and Paclitax NAB are most stable.
TABLE-US-00022 TABLE 22 Percentage of albumin in the form of dimers
on the nanoparticles. Sample Name N (number) Mean Margin of error
Min Max ABRAXANE .RTM. 66 17.8 1.1 6.7 25.7 Paclitax NAB 1 19.4 --
19.4 19.4 Albupax 1 16.5 -- 16.5 16.5 PacliALL 4 21.9 6.0 16.6 25.1
V5 FP 1 11.0 -- 11.0 11.0 V1 FP 1 11.3 -- 11.3 11.3 V2 FP 1 12.9 --
12.9 12.9 V3 FP 1 8.0 -- 8.0 8.0 V4 FP 1 9.8 -- 9.8 9.8 V5 IP 1
13.5 -- 13.5 13.5 V1 IP 1 10.9 -- 10.9 10.9 V2 IP 1 21.0 -- 21.0
21.0 V3 IP 1 11.6 -- 11.6 11.6 V4 IP 1 10.8 -- 10.8 10.8
A comparison of the percentage of albumin on the nanoparticles in
the form of dimers (Table 22) is illustrated in the bar graph of
FIG. 10. Amongst all the compositions, nab-paclitaxel sold under
the trademark ABRAXANE.RTM. and Paclitax NAB are most stable.
TABLE-US-00023 TABLE 23 Percentage of albumin in the form of
oligomers on the nanoparticles. Sample Name N (number) Mean Margin
of error Min Max ABRAXANE .RTM. 66 7.9 0.9 3.1 16.5 Paclitax NAB 1
12.3 -- 12.3 12.3 Albupax 1 9.1 -- 9.1 9.1 PacliALL 4 13.4 6.8 8.3
17.2 V5 FP 1 3.2 -- 3.2 3.2 V1 FP 1 3.3 -- 3.3 3.3 V2 FP 1 4.0 --
4.0 4.0 V3 FP 1 2.6 -- 2.6 2.6 V4 FP 1 2.7 -- 2.7 2.7 V5 IP 1 5.4
-- 5.4 5.4 V1 IP 1 3.0 -- 3.0 3.0 V2 IP 1 10.4 -- 10.4 10.4 V3 IP 1
2.7 -- 2.7 2.7 V4 IP 1 3.4 -- 3.4 3.4
A comparison of the percentage of albumin on the nanoparticles in
the form of oligomers (Table 23) is illustrated in the bar graph of
FIG. 11. Amongst all the compositions, nab-paclitaxel sold under
the trademark ABRAXANE.RTM. and Paclitax NAB are most stable.
TABLE-US-00024 TABLE 24 Percentage of albumin in the form of
polymers on the nanoparticles. Sample Name N (number) Mean Margin
of error Min Max ABRAXANE .RTM. 66 27.2 2.5 10.7 69.0 Paclitax NAB
1 23.5 -- 23.5 23.5 Albupax 1 18.8 -- 18.8 18.8 PacliALL 4 10.6 9.6
4.3 18.5 V5 FP 1 17.3 -- 17.3 17.3 V1 FP 1 12.7 -- 12.7 12.7 V2 FP
1 21.1 -- 21.1 21.1 V3 FP 1 33.2 -- 33.2 33.2 V4 FP 1 24.0 -- 24.0
24.0 V5 IP 1 15.0 -- 15.0 15.0 V1 IP 1 8.0 -- 8.0 8.0 V2 IP 1 16.8
-- 16.8 16.8 V3 IP 1 30.1 -- 30.1 30.1 V4 IP 1 15.9 -- 15.9
15.9
A comparison of the percentage of albumin on the nanoparticles in
the form of polymers (Table 24) is illustrated in the bar graph of
FIG. 12. Amongst all the compositions, nab-paclitaxel sold under
the trademark ABRAXANE.RTM. and Paclitax NAB are most stable.
Using the measurements of albumin in the form of monomers (M),
dimers (D), oligomers (O), and polymers (P) on the nanoparticles,
composition attributes were calculated as reported in Table 25.
TABLE-US-00025 TABLE 25 Summary of attributes calculated for
albumin forms on the nanoparticles in the compositions. (P + O)/ (P
+ O)/ Sample Name D/M O/M P/M M (M - D) M + D M - D M + O M + P M -
P D + O D + P O + P P/D O/D P/O ABRAXANE .RTM. 37.7 16.8 57.7 74.6
119.7 64.9 29.4 55.1 74.3 19.9 25.7 45.- 0 35.1 153.3 44.7 343.0
Paclitax NAB 43.3 27.5 52.5 79.9 140.9 64.2 25.4 57.1 68.3 21.3
31.7 42.9 - 35.8 121.1 63.4 191.1 Albupax 29.8 16.4 33.8 50.2 71.5
72.1 39.0 64.7 74.3 36.8 25.7 35.3 27.9 1- 13.5 55.1 205.9 PacliALL
40.5 24.8 19.6 44.4 74.6 76.0 32.2 67.5 64.8 43.5 35.3 32.5 24.0 -
48.5 61.2 79.3 V5 FP 16.0 4.7 25.3 30.0 35.7 79.5 57.6 71.7 85.8
51.2 14.2 28.3 20.5 157.- 8 29.5 535.6 V1 FP 15.5 4.5 17.4 21.9
25.9 84.0 61.5 76.0 85.5 60.1 14.5 24.0 16.0 112.- 7 29.0 389.3 V2
FP 20.9 6.5 34.1 40.6 51.4 74.8 49.0 65.9 83.1 40.8 17.0 34.1 25.2
163.- 5 31.1 525.9 V3 FP 14.2 4.5 58.9 63.4 73.9 64.3 48.3 58.9
89.5 23.2 10.5 41.1 35.7 416.- 1 32.1 1295.3 V4 FP 15.4 4.2 37.8
42.0 49.7 73.3 53.7 66.2 87.5 39.5 12.5 33.8 26.7 244.- 5 27.4
891.8 V5 IP 20.4 8.1 22.7 30.8 38.7 79.6 52.7 71.5 81.2 51.1 18.9
28.5 20.4 111.- 3 39.7 280.0 V1 IP 14.0 3.8 10.2 14.0 16.3 89.0
67.2 81.1 86.1 70.2 13.9 18.9 11.0 72.7- 27.4 265.0 V2 IP 40.4 20.0
32.3 52.3 87.6 72.9 31.0 62.3 68.7 35.2 31.3 37.7 27.1 80.- 0 49.5
161.4 V3 IP 20.8 4.8 54.0 58.8 74.3 67.2 44.1 58.4 85.8 25.6 14.3
41.6 32.8 260.- 1 23.3 1117.8 V4 IP 15.5 4.9 22.7 27.6 32.7 80.7
59.0 73.3 85.8 54.0 14.2 26.7 19.3 146.- 7 31.5 466.0
A comparison of the percentage of albumin on the nanoparticles in
the form of monomers (M) and dimers (D) (Table 25) is illustrated
in the bar graph of FIG. 13. Amongst all the compositions,
nab-paclitaxel sold under the trademark ABRAXANE.RTM. and Paclitax
NAB are most stable.
A comparison of the percentage of albumin on the nanoparticles in
the form of monomers (M) minus the percentage of albumin on the
nanoparticles in the form of dimers (D) (Table 25) is illustrated
in the bar graph of FIG. 14. Amongst all the compositions,
nab-paclitaxel sold under the trademark ABRAXANE.RTM. and Paclitax
NAB are most stable.
A comparison of the percentage of albumin on the nanoparticles in
the form of monomers (M) and oligomers (O) (Table 25) is
illustrated in the bar graph of FIG. 15. Amongst all the
compositions, nab-paclitaxel sold under the trademark ABRAXANE.RTM.
and Paclitax NAB are most stable.
A comparison of the percentage of albumin on the nanoparticles in
the form of monomers (M) and polymers (P) (Table 25) is illustrated
in the bar graph of FIG. 16. Amongst all the compositions,
nab-paclitaxel sold under the trademark ABRAXANE.RTM. and Paclitax
NAB are most stable.
A comparison of the percentage of albumin on the nanoparticles in
the form of monomers (M) minus the percentage of albumin on the
nanoparticles in the form of polymers (P) (Table 25) is illustrated
in the bar graph of FIG. 17. Amongst all the compositions,
nab-paclitaxel sold under the trademark ABRAXANE.RTM. and Paclitax
NAB are most stable.
A comparison of the percentage of albumin on the nanoparticles in
the form of dimers (D) and oligomers (O) (Table 25) is illustrated
in the bar graph of FIG. 18. Amongst all the compositions,
nab-paclitaxel sold under the trademark ABRAXANE.RTM. and Paclitax
NAB are most stable.
A comparison of the percentage of albumin on the nanoparticles in
the form of dimers (D) and polymers (P) (Table 25) is illustrated
in the bar graph of FIG. 19. Amongst all the compositions,
nab-paclitaxel sold under the trademark ABRAXANE.RTM. and Paclitax
NAB are most stable.
A comparison of the percentage of albumin on the nanoparticles in
the form of oligomers (O) and polymers (P) (Table 25) is
illustrated in the bar graph of FIG. 20. Amongst all the
compositions, nab-paclitaxel sold under the trademark ABRAXANE.RTM.
and Paclitax NAB are most stable.
A comparison of the ratio (as reported as a percentage) of the
percentage of albumin on the nanoparticles in the form of dimers
(D) divided by the percentage of albumin on the nanoparticles in
the form of monomers (M) (Table 25) is illustrated in the bar graph
of FIG. 21. Amongst all the compositions, nab-paclitaxel sold under
the trademark ABRAXANE.RTM. and Paclitax NAB are most stable.
A comparison of the ratio (as reported as a percentage) of the
percentage of albumin on the nanoparticles in the form of oligomers
(O) divided by the percentage of albumin on the nanoparticles in
the form of monomers (M) (Table 25) is illustrated in the bar graph
of FIG. 22. Amongst all the compositions, nab-paclitaxel sold under
the trademark ABRAXANE.RTM. and Paclitax NAB are most stable.
A comparison of the ratio (as reported as a percentage) of the
percentage of albumin on the nanoparticles in the form of polymers
(P) divided by the percentage of albumin on the nanoparticles in
the form of monomers (M) (Table 25) is illustrated in the bar graph
of FIG. 23. Amongst all the compositions, nab-paclitaxel sold under
the trademark ABRAXANE.RTM. and Paclitax NAB are most stable.
A comparison of the ratio (as reported as a percentage) of the
percentage of albumin on the nanoparticles in the form of polymers
(P) and oligomers (O) divided by the percentage of albumin on the
nanoparticles in the form of monomers (M) (Table 25) is illustrated
in the bar graph of FIG. 24. Amongst all the compositions,
nab-paclitaxel sold under the trademark ABRAXANE.RTM. and Paclitax
NAB are most stable.
A comparison of the ratio (as reported as a percentage) of the
percentage of albumin on the nanoparticles in the form of polymers
(P) and oligomers (O) divided by the percentage of albumin on the
nanoparticles in the form of monomers (M) minus dimers (D) (Table
25) is illustrated in the bar graph of FIG. 25. Amongst all the
compositions, nab-paclitaxel sold under the trademark ABRAXANE.RTM.
and Paclitax NAB are most stable.
TABLE-US-00026 TABLE 26 Solubility (.mu.g/ml) and dissolution
kinetics of the composition immediately after reconstitution.
Dissolution Sample Name N (number) Mean Margin of error Min Max
kinetics comment ABRAXANE .RTM. 11 66.1 5.3 53.5 82.2 Normal
Paclitax NAB 1 63.3 -- 63.3 63.3 Normal Albupax 1 40.0 -- 40.0 40.0
-- PacliALL 3 85.0 21.2 76.0 93.0 Normal V5 FP 3 80.0 39.8 62.3
93.5 Normal V1 FP 3 66.2 22.4 58.0 75.9 Normal V2 FP 2 70.9 183.0
56.5 85.3 None V3 FP 3 74.8 13.6 68.8 79.5 Normal V4 FP 3 69.1 22.1
59.8 77.5 Slower than normal
TABLE-US-00027 TABLE 27 Degree of sedimentation (stability) based
on visual observation of the composition (1 indicates no
sedimentation) immediately after reconstitution. Sample Name N
(number) Mean* Margin of error Min Max ABRAXANE .RTM. 17 1 0 1 1
Paclitax NAB 2 1 0 1 1 Albupax -- -- -- -- -- PacliALL 3 1 0 1 1 V5
FP 3 1 0 1 1 V1 FP 3 1 0 1 1 V2 FP 3 1 0 1 1 V3 FP 3 1 0 1 1 V4 FP
3 1 0 1 1 *The degree of sedimentation recited as: 1 - No visible
sedimentation (NVS); 2 - Streaming with NVS; 3 - Very slight
sedimentation; 4 - Slight sedimentation; 5 - Sedimentation; 6 -
Phase separation.
*The degree of sedimentation recited as: 1--No visible
sedimentation (NVS); 2--Streaming with NVS; 3--Very slight
sedimentation; 4--Slight sedimentation; 5--Sedimentation; 6--Phase
separation.
TABLE-US-00028 TABLE 28 Percentage of paclitaxel recovered after
filtration through a 0.2-.mu.m syringe filter immediately after
reconstitution. Sample Name N (number) Mean Margin of error Min Max
ABRAXANE .RTM. 21 98.2 1.4 93.3 106.2 Paclitax NAB 2 104.0 76.2 98
110 Albupax -- -- -- -- -- PacliALL 5 43.0 24.1 27 72 V5 FP 3 95.7
8.7 92 99 V1 FP 3 92.3 18.0 84 97 V2 FP 2 92.0 63.5 87 97 V3 FP 3
97.0 7.5 94 100 V4 FP 3 99.0 13.1 95 105
TABLE-US-00029 TABLE 29 Solubility (.mu.g/ml) and dissolution
kinetics of the composition after storage of the reconstituted
suspension for 24 hours at 40.degree. C. Dissolution N Margin
Solubility kinetics Sample Name (number) Mean of error Min Max
comment comment ABRAXANE .RTM. 12 66.8 7.5 54.0 96.8 Regular Normal
Paclitax NAB 1 44.9 -- 44.9 44.9 Low solubility Normal Albupax --
-- -- -- -- -- Normal PacliALL 3 46.9 101.1 0.0 73.5
Insoluble/Regular -- V5 FP 2 74.1 2.5 73.9 74.3 ~Regular Normal V1
FP 2 0.0 0.0 0.0 0.0 Insoluble None V2 FP 1 0.0 -- 0.0 0.0
Insoluble None V3 FP 2 0.0 0.0 0.0 0.0 Insoluble None V4 FP 2 0.0
0.0 0.0 0.0 Insoluble None
TABLE-US-00030 TABLE 30 Degree of sedimentation (stability) based
on visual observation (1 indicates no sedimentation) and
crystallinity (1 indicates presence of crystalline paclitaxel) of
the composition after storage of the reconstituted suspension for
24 hours at 40.degree. C. N Margin Crystallinity Sample Name
(number) Mean* of error Min Max Crystallinity value ABRAXANE .RTM.
17 1 0 1 1 No Birefringence 0 Paclitax NAB 2 1 0 1 1 No
Birefringence 0 Albupax 1 5 -- 5 5 Birefringence 1 PacliALL 5 5 0 5
5 Birefringence 1 V5 FP 2 5 0 5 5 Birefringence 1 V1 FP 2 5.5 6.4 5
6 Birefringence 1 V2 FP 1 6 0 6 6 Birefringence 1 V3 FP 2 6 0 6 6
Birefringence 1 V4 FP 2 6 0 6 6 Birefringence 1 *The degree of
sedimentation recited as: 1 - No visible sedimentation (NVS); 2 -
Streaming with NVS; 3 - Very slight sedimentation; 4 - Slight
sedimentation; 5 - Sedimentation; 6 - Phase separation.
TABLE-US-00031 TABLE 31 Degree of sedimentation (stability) based
on visual observation (1 indicates no sedimentation) and
crystallinity (1 indicates presence of crystalline paclitaxel) of
the composition after storage of the reconstituted suspension at
40.degree. C. for 16 hours. Crystallinity Sample Name Mean value V5
FP 2.7 0 V1 FP 5 1 V2 FP 5.5 1 V3 FP 5.3 0.66 V4 FP 5 0.66
TABLE-US-00032 TABLE 32 Percentage of paclitaxel recovered after
filtration through a 0.2-.mu.m syringe filter after storage of the
reconstituted suspension at 40.degree. C. for 16 hours. Sample Name
Mean V5 FP 96.5 V1 FP 59.2 V2 FP 22.6 V3 FP 50.1 V4 FP 36.2
TABLE-US-00033 TABLE 33 Degree of sedimentation (stability) based
on visual observation (1 indicates no sedimentation) and
crystallinity (1 indicates presence of crystalline paclitaxel) of
the composition after storage of the reconstituted suspension at
5.degree. C. for 8 hours followed by storage at 25.degree. C. for 8
hours. Sample Name Mean Crystallinity value V5 FP 1 0.333 V1 FP
1.33 1 V2 FP 1.5 0 V3 FP 1.33 0 V4 FP 1 0
TABLE-US-00034 TABLE 34 Percentage of paclitaxel recovered after
filtration through a 0.2-.mu.m syringe filter after storage of the
reconstituted suspension at 5.degree. C. for 8 hours followed by
storage at 25.degree. C. for 8 hours. Sample Name Mean V5 FP 98.7
V1 FP 72.7 V2 FP 64.0 V3 FP 91.0 V4 FP 92.0
TABLE-US-00035 TABLE 35 Percentage of paclitaxel recovered after
filtration through a 0.2-.mu.m syringe filter after storage of the
reconstituted suspension for 24 hours at 40.degree. C. N Margin
Sample Name (number) Mean of error Min Max ABRAXANE .RTM. 18 96.4
1.7 92.2 106.7 Paclitax NAB -- -- -- -- -- Albupax 1 61 -- 61 61
PacliALL 1 69 -- 69 69 V5 FP 2 86.3 36.8 83.4 89.2 V1 FP 2 13.5
170.9 0.0 26.9 V2 FP 1 0.0 -- 0.0 0.0 V3 FP 2 9.7 91.5 2.5 16.9 V4
FP 2 0.0 0.0 0.0 0.0
TABLE-US-00036 TABLE 36 D.sub.V4,3 (nm) measured after storage of
the reconstituted suspension for 24 hours at 40.degree. C. N Margin
Sample Name (number) Mean of error Min Max ABRAXANE .RTM. 20 155.8
3.6 143.5 169.1 Paclitax NAB 2 118.4 48.9 114.5 122.2 Albupax 1
150.9 -- 150.9 150.9 PacliALL 5 1052.5 3531.3 231.5 2694 V5 FP 2
172.9 90.8 165.7 180 V1 FP 2 2556.5 7096.4 1998 3115 V2 FP 1 2475.0
-- 2475 2475 V3 FP 2 2408.0 5539.9 1972 2844 V4 FP 2 2431.0 7814.3
1816 3046
TABLE-US-00037 TABLE 37 Z average (nm) measured after storage of
the reconstituted suspension for 24 hours at 40.degree. C. N Margin
Sample Name (number) Mean of error Min Max ABRAXANE .RTM. 20 147.7
2.2 139.5 155.8 Paclitax NAB 2 121.7 37.4 118.8 124.7 Albupax 1
157.7 -- 157.7 157.7 PacliALL 5 1411.3 2634.2 183.4 5098 V5 FP 2
156.9 60.3 152.2 161.7 V1 FP 2 2361.8 25414.2 361.7 4362 V2 FP 1
3435.0 -- 3435 3435 V3 FP 2 692.0 4459.2 341.1 1043 V4 FP 2 5338.5
946.6 5264 5413
TABLE-US-00038 TABLE 38 Polydispersity index (PDI) measured after
storage of the reconstituted suspension for 24 hours at 40.degree.
C. N Margin Sample Name (number) Mean of error Min Max ABRAXANE
.RTM. 20 0.11555 0.007141 0.092 0.156 Paclitax NAB 2 0.1015
0.069884 0.096 0.107 Albupax 1 0.214 -- 0.214 0.214 PacliALL 5
0.3806 0.353505 0.178 0.862 V5 FP 2 0.141 0.1270619 0.131 0.151 V1
FP 2 0.6135 2.12829 0.446 0.781 V2 FP 1 0.315 -- 0.315 0.315 V3 FP
2 0.8545 0.603542 0.807 0.902 V4 FP 2 0.368 1.4358 0.255 0.481
TABLE-US-00039 TABLE 39 DV5 (nm) measured after storage of the
reconstituted suspension for 24 hours at 40.degree. C. N Margin
Sample Name (number) Mean of error Min Max ABRAXANE .RTM. 20 76.2
2.4 65.2 84.7 Paclitax NAB 2 63.2 6.3 62.7 63.7 Albupax 1 86.3 --
86.3 86.3 PacliALL 5 611.2 944.5 73.8 1760 V5 FP 2 73.8 94.0 66.4
81.2 V1 FP 2 834 8716.4 148 1520 V2 FP 1 1500.0 -- 1500 1500 V3 FP
2 172.5 209.6 156 189 V4 FP 2 1715.0 4637.7 1350 2080
TABLE-US-00040 TABLE 40 DV50 (nm) measured after storage of the
reconstituted suspension for 24 hours at 40.degree. C. N Margin
Sample Name (number) Mean of error Min Max ABRAXANE .RTM. 20 140.2
4.138 124 154 Paclitax NAB 2 107 38.11875 104 110 Albupax 1 142 --
142 142 PacliALL 5 1075.2 1466.9505 200 2500 V5 FP 2 151 139.768
140 162 V1 FP 2 2795 10355.52 1980 3610 V2 FP 1 2337 -- 2337 2337
V3 FP 2 2445 6035.44 1970 2920 V4 FP 2 2390 7369.59 1810 2970
TABLE-US-00041 TABLE 41 DV95 (nm) measured after storage of the
reconstituted suspension for 24 hours at 40.degree. C. N Margin
Sample Name (number) Mean of error Min Max ABRAXANE .RTM. 20 290.9
7.695 267 336 Paclitax NAB 2 214.5 133.41495 204 225 Albupax 1 249
-- 249 249 PacliALL 5 3062.4 2951.1995 496 5450 V5 FP 2 346 63.531
341 351 V1 FP 2 4305 21664 2600 6010 V2 FP 1 3917 -- 3917 3917 V3
FP 2 4265 15565.1 3040 5490 V4 FP 2 3345 13023.82 2320 4370
TABLE-US-00042 TABLE 42 (DV90 - DV10)/DV50 measured after storage
of the reconstituted suspension for 24 hours at 40.degree. C. N
Margin Sample Name (number) Mean of error Min Max ABRAXANE .RTM. 20
1.20616 0.055545 1.03684 1.46048 Paclitax NAB 2 1.08277 0.611737
1.03462 1.13091 Albupax 1 0.88803 -- 0.88803 0.88803 PacliALL 5
3.76422 6.98718 0.736 13.8008 V5 FP 2 1.43079 2.077525 1.26728
1.59429 V1 FP 2 0.955875 6.498385 0.44444 1.46731 V2 FP 1 0.82713
-- 0.82713 0.82713 V3 FP 2 1.17052 6.101 0.69036 1.65068 V4 FP 2
0.517895 1.0347265 0.43646 0.59933
Example 15. Characterization of a Composition Comprising
Nanoparticles Comprising Rapamycin and Albumin
A composition comprising nanoparticles comprising rapamycin and
albumin was prepared and reconstituted as described above to form a
reconstituted suspension. A sample of the reconstituted suspension
was ultracentrifuged as described above to form a supernatant and a
pellet containing the nanoparticles. The following concentrations
were determined using the methods described above for quantifying
rapamycin (RP-HPLC) and albumin (HPLC size-exclusion
chromatography): rapamycin in the supernatant, rapamycin in the
pellet, rapamycin in the reconstituted suspension, human albumin
(HA) in the supernatant, HA in the pellet, HA in the reconstituted
suspension, HA in a 4% HA stock solution, and HA in 4% HA system
suitability. See Table 43. The percentage of albumin monomers,
dimers, oligomers, and polymers in the following samples from the
ultracentrifugation study was determined as described above: the
supernatant, the pellet, the reconstituted suspension, a 4% HA
stock solution, and 4% HA system suitability. See Table 44.
TABLE-US-00043 TABLE 43 Assay Assay Results Results Sample Sample
ID (mg/mL) (Average) 1 Rapamycin in Supernatant 0.054 0.055 2 0.054
3 0.057 4 Rapamycin in Pellet 4.79 4.765 5 4.75 6 4.76 7 Rapamycin
in Reconstituted 5.09 5.085 8 Suspension 5.09 9 5.07 10 HA in
Supernatant 43.52 43.515 11 43.48 12 43.54 13 HA in Reconstituted
Suspension 46.07 46.008 14 46.05 15 45.91 16 4% HA Stock Solution
43.72 43.718 17 4% HA System Suitability 41.06 41.048 18 41.06 19
41.00 20 41.06 21 41.06 22 41.03 23 41.06 24 HA in Pellet 1.65
1.651 25 1.65 26 1.65
TABLE-US-00044 TABLE 44 Unnamed Monomer Dimer Oligomer Polymer 4%
HA Stock Solution 95.2 1.9 0.3 2.5 4% HA System suitability 97.2
1.9 0.3 0.7 HA in Reconstituted 82.0 12.9 3.7 1.4 Suspension 82.1
12.9 3.7 1.3 81.8 13.0 3.8 1.3 HA Supernatant 82.5 13.1 3.9 0.6
82.9 12.9 3.6 0.6 82.7 13.0 3.8 0.5 HA Pellet 39.0 21.4 12.5 27.0
37.2 21.6 13.1 28.1 36.0 21.9 13.5 28.6
Mass balance calculations were carried out for rapamycin (see Table
45) and HA (see Table 46), showing 5.2% and 2.5% differences for
rapamycin and HA, respectively, for their concentrations in the
reconstituted suspension compared to their concentrations in the
ultracentrifugation fractions. As shown in Table 47, the ratios
(w/w) for rapamycin and HA in the supernatant versus the pellet
fractions was 98.9:1.1 and 96.3:3.7, respectively, and the ratio
(w/w) of rapamycin to HA in the pellet fraction, was 74:26.
TABLE-US-00045 TABLE 45 Concentration of Concentration
Concentration of Rapamycin in of Rapamycin Rapamycin in
Reconstituted in Pellet Supernatant Suspension (mg/mL) (mg/mL)
(mg/mL) % Difference 4.765 0.055 5.1 5.2
TABLE-US-00046 TABLE 46 Concentration of Concentration HA in of HA
in Concentration of Reconstituted Pellet HA in Supernatant
Suspension (mg/mL) (mg/mL) (mg/mL) % Difference 1.651 45.5 46.0
2.5
TABLE-US-00047 TABLE 47 Rap HA Rap:HA (Supernatant):Rap
(Supernatant):HA (Pellet) (Pellet) (w/w) (Pellet) (w/w) (w/w)
98.9:1.1 96.3:3.7 74:26
* * * * *
References